Deletion of miR-130b/301b cluster promotes macrophage efferocytosis and resolution of adipose tissue inflammation

Lei Yang; Tuerdiguli Abuduyimiti; Yibo Xi; Gianfilippo Coppola; Chen Liu; Shaoning Jiang

doi:10.1016/j.isci.2026.115307

. 2026 Mar 10;29(4):115307. doi: 10.1016/j.isci.2026.115307

Deletion of miR-130b/301b cluster promotes macrophage efferocytosis and resolution of adipose tissue inflammation

Lei Yang ¹, Tuerdiguli Abuduyimiti ¹, Yibo Xi ¹, Gianfilippo Coppola ¹, Chen Liu ¹, Shaoning Jiang ^1,^2,^∗

PMCID: PMC13049439 PMID: 41940338

Summary

Macrophage efferocytosis, the clearance of apoptotic cells, is essential for tissue homeostasis and preventing inflammation. Impaired efferocytosis contributes to chronic inflammatory conditions, including obesity. However, its key regulators remain unclear. MicroRNA-130b (miR-130b) is increased in adipose tissue macrophages of individuals with obesity. Here, we found that miR-130b was enriched in bone marrow of mice, and its expression in bone marrow-derived macrophages was suppressed by IL-4 and by apoptotic cell uptake. Deletion of the miR-130b and its cluster member miR-301b enhanced macrophage efferocytosis in vitro and apoptotic cell clearance in vivo, accompanied by increased mitochondrial respiration and anti-inflammatory polarization. In high fat diet-fed mice, global deletion of miR-130b/301b reduced inflammatory gene expression in adipose tissues. Mechanistically, miR-130b suppressed PPARγ and PGC-1α, regulators of mitochondrial metabolism and inflammation, and miR-130b/301b deletion increased CX3CR1, a receptor for apoptotic cell “find me” signals. Together, miR-130b/301b deletion promotes macrophage efferocytosis and resolves adipose tissue inflammation.

Subject areas: molecular biology, immunology, cell biology

Graphical abstract

Highlights

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Macrophage expression of miR-130b/301b is suppressed by phagocytosis of apoptotic cells
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miR-130b/301b deletion promoted macrophage migration and efferocytosis
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miR-130b/301b deletion enhanced mitochondrial respiration
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miR-130b/301b deletion attenuated HFD-induced adipose tissue inflammation

Molecular biology; Immunology; Cell biology

Introduction

Macrophage phagocytosis of apoptotic cells (ACs), a process known as efferocytosis, plays a crucial role in maintaining tissue homeostasis and resolution of inflammation.¹^,²^,³ Under normal conditions in an adult, billions of cells undergo apoptosis daily, a regulated form of programmed cell death where the dead cells are rapidly removed by efferocytosis without triggering the inflammatory response.⁴^,⁵ In contrast, defective removal of ACs due to impaired macrophage efferocytosis can result in necrosis of ACs, leading to release of cytoplasmic contents, including damage-associated molecular patterns (DAMPs), which induce inflammation and disrupt tissue homeostasis.⁴ Impaired efferocytosis has been implicated in various pathological conditions, including obesity and the associated chronic inflammation.⁶ In obesity, macrophage lipid overload and defective efferocytosis contribute to local and systemic chronic inflammation and metabolic dysfunction.⁷^,⁸^,⁹ A better understanding of the regulatory mechanisms of efferocytosis is crucial for developing targeted therapies to promote the resolution of inflammation.

Efferocytosis is a multistep, fine-tuned process that involves the recruitment of macrophages, engulfment of ACs, and degradation of the cellular contents. During apoptosis, cells release a variety of “find-me” signals to attract macrophages and promote a pro-resolving milieu, such as lysophosphatidylcholine (LPC), sphingosine-1-phosphate (S1P), nucleotides, and CX3CL1.¹ In addition, ACs display “eat-me” signals on their cell surface, such as phosphatidylserine (PS) and calreticulin.¹⁰ Those signals are recognized by macrophages directly or indirectly via receptors and bridge molecules, such as the PS receptor, growth arrest-specific protein 6 (CAS6), integrins, tyrosine-kinase-activated receptor (Mer-TK), and CX3CL1 receptor CX3CR1.²^,¹¹ Upon recognition of a dying cell, the phagocyte initiates cytoskeletal remodeling that allows for the formation of the phagosome.¹¹ With engulfment of ACs, macrophages are subjected to a large metabolic load, which results in elevated mitochondrial respiration, promoting macrophage anti-inflammatory reprogramming in part through lipid-activated transcription factors, such as peroxisome proliferator-activated receptor (PPAR) gamma and retinoid × receptor (RXR).¹⁰^,¹²^,¹³ Activation of these transcription factors promotes upregulation of phagocytic receptors and bridging molecules, and anti-inflammatory mediators.¹¹ Mitochondria play a crucial role in efferocytosis and efferocytosis-associated resolution of inflammation, including cellular energy production, calcium regulation, and degradation of ACs.¹²^,¹⁴^,¹⁵ Despite increased identification of numerous factors linked to efferocytosis in physiological and pathological conditions,² key regulators and the precise mechanisms of these processes remain to be uncovered.

MicroRNAs are small noncoding RNAs that influence gene expression post-transcriptionally in a cell type-specific manner.¹⁶ Accumulating evidence suggests a pivotal role of microRNAs in regulating macrophage differentiation and function.¹⁷ The microRNA cluster, miR-130b/301b, includes miR-130b and miR-301b, which share the same seed sequence and are located next to each other in the genome. Previous studies by our group and others have identified miR-130b and/or miR-301b to be dysregulated in human obesity, dyslipidemia, and gestational diabetes.¹⁸^,¹⁹^,²⁰^,²¹ Notably, expression of miR-130b is increased in both adipocytes and adipose tissue macrophages (ATMs) of individuals with obesity.¹⁹ We have previously defined a role of miR-130b/301b in inhibiting beige adipogenesis in obesity.²² In the present study, we further investigated the regulation and function of miR-130b/301b in macrophages.

Results

Decreased miR-130b expression following differentiation of BMDMs and in response to IL-4 anti-inflammatory stimulation

Tissue distribution analysis of miR-130b-3p in the mouse showed a highly enriched miR-130b-3p expression in bone marrow compared to other metabolically active tissues, including liver, muscle, and fat tissues (Figure 1A). Differentiation of bone marrow progenitor cells into bone marrow-derived macrophages (BMDMs) using macrophage colony stimulating factor (M-CSF) resulted in significant suppression of miR-130b expression (Figure 1B). To further explore the factors regulating miR-130b expression, BMDMs were treated with either anti-inflammatory factor IL-4 or pro-inflammatory activators TNFα and IL-1β. As the results shown in Figure 1C, expression of miR-130b-3p was suppressed by IL-4 treatment, indicating a negative regulation of miR-130b-3p expression by macrophage anti-inflammatory activation. On the other hand, TNFα or IL-1β treatment significantly increased expression of both miR-130b-3p and miR-301b-3p (Figure 1D).

Regulation of miR-130b expression in bone marrow-derived macrophages

(A) iWAT, eWAT, BAT, quadriceps muscle, liver, and bone marrow tissues were collected from C57BL/6J male mice (5–7 weeks of age, normal diet). Total RNAs were extracted and relative expression of miR-130b-3p in indicated tissues were measured by qPCR, and fold differences to iWAT were shown (n = 4 mice).

(B) Expression of miR-130b-3p in bone marrow progenitor cells at day 0, 3, and 5 following bone marrow macrophages differentiation using M-CSF (30 μg/mL, n = 3 cell samples).

(C) Expression of miR-130b-3p in bone marrow-derived macrophages (BMDM) in response to IL-4 (5 ng/mL) treatment for 5 h (n = 3 cell samples); (D) Expression of miR-130b-3p and miR-301b in BMDMs in response to TNFα (5 ng/mL) or IL-1β (1 ng/mL) treatment for 2 days (n = 4 cell samples).

(E and F) Bone marrow-derived macrophages were incubated with apoptotic thymocytes (6 × 10⁵ cells) for 6 h, followed by qPCR analysis of *Arg-1* and *Il-10* expression (E) and miR-130b-3p and miR-310b-3p expression (F). n = 3 cell samples in each group. All bar graphs were presented as mean ± SD, ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001 by t test compared between control and treatment groups.

Decreased miR-130b and miR-301b in BMDMs following phagocytosis of ACs

Since efferocytosis is associated with an anti-inflammatory response, we next examined the regulation of miR-130b/miR-301b expression following macrophage efferocytosis. Apoptotic thymocytes were added to primary macrophages to induce efferocytosis, and after 6 h, expression of miR-130b/301b and anti-inflammatory genes were measured. Engulfment of ACs by bone marrow-derived macrophages resulted in a significant increase in the expression of anti-inflammatory genes Arg-1 and IL-10 (Figure 1E), concomitant with decreased expression of miR-130b-3p, as well as the cluster member miR-301b-3p (Figure 1F).

Deletion of miR-130b/301b cluster enhanced macrophage efferocytosis in vitro and in vivo

We next determined the roles of the miR-130b/301b cluster in macrophage function of efferocytosis. Peritoneal macrophages from mice lacking both miR-130b and miR-301b (miR-130b/301b cluster knockout [KO] mice) or wild-type (WT) controls were incubated with CFSE (Carboxyfluorescein diacetate N-succinimidyl ester)-labeled apoptotic thymocytes for 2 h. As gated by flow cytometry analysis, the cells with CFSE positive while negative for CD90.1 (the cell surface marker of thymocytes) represent macrophages with engulfment of apoptotic thymocytes (Figure 2A). Macrophages with CD90.1 positive are cells with thymocytes attached to the cell surface that are not engulfed (Figure 2A). As results of flow cytometry shown in Figure 2A, the percentage of macrophages with engulfment of ACs and the intensity were significantly higher in miR-130b/301b KO macrophages, suggesting deletion of the miR-130b/301b cluster enhanced macrophage efferocytosis in vitro.

Deletion of miRNA-130b and miR-301b-enhanced macrophage efferocytosis *in vitro* and *in vivo*

(A) Peritoneal macrophages from miR-130b/301b KO or WT control mice were incubated with CFSE-labeled apoptotic thymocytes. After 2 h, the cells were labeled with BV421-conjugated antibody targeting CD90.1 and subjected to flow cytometry. Left shows gating of the cells for apoptotic cells engulfed (CFSE+ CD90.1-) and attached (CFSE+ CD90.1+) macrophages. Bar graphs show percentage and intensity of CFSE positive and CD90.1 negative cells. n = 4 cell samples.

(B) Peritoneal macrophages from miR-130b/301b KO or WT control mice were incubated with or without GFP-expressing apoptotic Hepa1-6 cells. After 1 h, the cells were labeled with F4/80 antibody and subjected to flow cytometry. Gating strategies and the percentage of GFP (FITC) positive cells in population of macrophages (F4/80) were shown. n = 3 cell samples.

(C and D) miR-130b/301b KO or WT control mice were subjected to dexamethasone (Dex) (10 mg/kg, intraperitoneally) or PBS (control) for 18 h.

(C) The thymus tissues were sectioned and subjected to TUNEL staining and representative images and quantified intensity were shown.

(D) Representative thymus images and the ratio of the weight of thymuses to body weight of mice that received dexamethasone. n = 3–4 mice in each group from two independent repeats. All bar graphs were presented as mean ± SD, ∗p < 0.5, ∗∗∗p < 0.001(t test for A, B, D, and ANOVA for C).

The effect of miR-130b/301b on macrophage efferocytosis was further confirmed by using ACs from a different source. A mouse hepatoma cell line, Hepa1-6 cells with GFP stable expression, were induced to apoptosis, and then incubated with peritoneal macrophages from mice lacking miR-130b/301b or WT controls. As results shown in Figure 2B, macrophages lacking miR-130b/301b exhibited higher phagocytosis of apoptotic Hepa1-6 cells compared to WT cells as assessed by counting the percentage of GFP-positive cells in F4/80⁺ macrophages via flow cytometry.

To further examine the role of miR-130b/301b in the clearance of ACs in vivo, we utilized an established method of dexamethasone (Dex)-induced apoptosis of thymocytes. Dex treatment resulted in increased ACs as measured by TUNEL staining (Figure 2C). Compared to WT controls, global deletion of miR-130b/301b significantly decreased apoptotic thymocytes (Figure 2C), and the size of thymus after Dex treatment was smaller in KO mice (Figure 2D), suggesting that deletion of miR-130b/301b enhanced clearance of ACs in vivo.

miR-130b/301b regulates macrophage inflammatory polarization

The impacts of miR-130b/301b deletion in macrophage polarization were next determined in the context of efferocytosis. Peritoneal macrophages from miR-130b/301b KO and WT mice were incubated with apoptotic thymocytes or vehicle control for 6 h. The expression of pro-inflammatory gene Mcp-1 and anti-inflammatory gene Ym-1 was measured. As results shown in Figure 3A, with or without engulfment of ACs, expression of Mcp-1 was significantly lower in miR-130b/301b-deleted macrophages compared to WT macrophages. In contrast, the expression of Ym-1 was higher in macrophages with miR-130b/301b deletion (Figure 3A). These results suggest a role of miR-130b/301b deletion in inducing macrophage anti-inflammatory polarization.

Role of miR-130b/301b in regulating macrophage polarization

(A) Peritoneal macrophages from miR-130b/301b KO mice or wild-type controls were incubated with apoptotic thymocytes. mRNA expression of *Mcp-1* and *Ym-1* in peritoneal macrophages following incubation with (engulfment) or without (con) apoptotic thymocytes for 6 h (n = 4 cell samples).

(B) Bone marrow-derived macrophages were transfected with the mimics of miR-130b-3p (50 nmol/L) and miR-301b-3p (50 nmol/L) or equal amount of a scrambled negative control for three days followed by activation with palmitate (0.5 mM) overnight. Levels of *Mcp-1* and *Tnf-α* mRNAs were measured by real-time PCR and normalized with TBP (n = 3 cell samples). All bar graphs were presented as mean ± SD. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001 (t test).

The specific roles of miR-130b and miR-301b overexpression in regulating macrophage inflammation were further determined. Overexpression of miR-130b mimic or miR-301b mimic in macrophages enhanced expression of proinflammatory factors Mcp-1 and Tnf-α following palmitate treatment (Figure 3B), consistent with a proinflammatory role of miR-130b and miR-301b.

Deletion of miR-130b/301b enhanced mitochondrial respiration

Mitochondria play an important role in the process of efferocytosis, and macrophage anti-inflammatory responses in macrophages rely on mitochondrial oxidative phosphorylation for energy production. We next determined the effects of miR-130b/301b deletion on macrophage mitochondrial function with or without efferocytosis. Seahorse Extracellular Flux analyzer was used to measure oxygen consumption rates (OCRs) over time at basal level and following application of oligomycin, FCCP (Carbonylcyanide-p-trifluoromethoxyphenylhydrazone), and antimycin A (AA)/Rotenone (ROT) as shown in Figure 4A. Efferocytosis of ACs resulted in increased basal and ATP-linked OCRs (Figure 4B). Compared to macrophages from WT mice, macrophages with deletion of the miR-130b/301b cluster exhibited increased basal and ATP-linked OCRs, with efferocytosis of ACs (Figure 4B), suggesting enhanced mitochondrial oxidative phosphorylation.

Effects of deletion of miR-130b/301b on macrophage mitochondrial respiration and pro-inflammatory polarization

Peritoneal macrophages from miR-130b/301b KO or WT control mice with or without incubation of apoptotic cells (AC, 1 × 10⁶, overnight) were subjected to seahorse mitochondrial respiration measurement.

(A) Oxygen consumption rate (OCR) was measured over time and after stress treatment of oligomycin (oligo, 1 mmol/L), FCCP (1 mmol/L), and rotenone and antimycin A (R/A) (0.5 mmol/L).

(B) Indices of baseline OCRandATP-linked OCR were calculated according to altered OCR following the treatment. Mean ± SEM, n = 5–7 cell samples. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 (ANOVA).

miR-130b significantly suppressed protein expression of PPARγ and PGC-1α

Previous studies have validated that miR-130b directly targets PPARγ²³^,²⁴ and PGC-1α²¹ (Figure 5A), which are key regulators of mitochondrial metabolism and anti-inflammatory response in macrophages. We next determined the regulation of PPARγ and PGC-1α protein expression by miR-130b/301b. In bone marrow macrophages, overexpression of miR-130b mimic or miR-301b resulted in significant decrease in PPARγ and PGC-1α protein expression (Figure 5B). In contrast, PPARγ and PGC-1α proteins were increased in macrophages with deletion of miR-130b/301b cluster (Figure 5C). Those findings suggest an important role of miR-130b and miR-301b in suppressing both PPARγ and PGC-1α protein expression in macrophages.

miR-130b inhibits macrophage expression of PGC-1α and PPARγ protein

(A) Sequences of PPAR and PGC-1 3′-UTR targeted by miR-130b-3p and miR-301b-3p.

(B) Bone marrow-derived macrophages (BMDMs) were transfected with miR-130b-3p mimics, miR-301b-3p mimics, or negative control (con). Total proteins were subjected for WB analysis at 2 days after transfection. Blots and quantifications of PGC-1α and PPARγ proteins (normalized to GAPDH of the same blot) were shown.

(C) BMDMs were isolated from WT and miR-130b/301B knockout mice (KO) and incubated with apoptotic thymocytes. After engulfment for 6 h, the cells were replaced with fresh medium and cultured for 2 days before collection for WB analysis. WB blots and quantifications of PGC-1α and PPARγ proteins (normalized to βactin from same blot) were shown. Bar graphs were shown as mean ± SD, n = 3–4 cell samples, ∗p < 0.05; ∗∗p < 0.01 (t test for B; ANOVA for C).

Differential gene expression profiles and pathways in macrophages with miR-130b/301b deletion

To better understand the profiles of genes and signaling pathways regulated by miR-130b/301b in macrophages, unbiased RNA sequencing (RNA-seq) analysis was performed on bone marrow-derived macrophages from miR-130b/301b KO and WT controls. Compared to WT control macrophages, deletion of miR-130b/301b resulted in upregulation of 219 genes and downregulation of 349 genes significantly (Figure 6A). Heatmap showed top 25 upregulated and downregulated genes (Figure 6B). Gene pathway enrichment analysis revealed that macrophage miR-130b/301b deletion upregulated genes involved in cellular energy and immune signaling pathways, including Adenyl nucleotide binding and immune cell receptor binding signaling (Figure 6C, list of genes in Table S2). By contrast, downregulated genes were associated with signaling pathways involved in growth factor binding and degradation of the extracellular matrix (Figure 6C, list of genes in Table S2). The randomly selected up- and down-regulated genes hit in the top pathways were validated by qPCR, including ATPase type 10D (Atp10d), the CX3CL1 receptor (Cx3cr1), and matrix metalloproteinases (Mmps), etc. (Figure 6D). Of note, Cx3cr1 mRNA (Figure 6E) expression was increased in response to engulfment of apoptotic thymocytes and further increased in macrophages with deletion of miR-130b/301b. Similarly, protein abundance of CX3CR1 was significantly higher in macrophages with deletion of miR-130b/301b following engulfment of ACs (Figure 6F).

RNA-seq analysis of BMDM from miR-130b/301b KO and WT control mice

(A) Volcano plot of RNA-seq analysis of differentially expressed genes between bone marrow-derived macrophages (BMDMs) from miR-130b/301b KO and WT mice. The x axis represents the log2 fold-change, and the y axis represents the –log10 p value. The significant genes are depicted in colors as indicated in figure legend. n = 3 mice in each group.

(B) Heatmap of top 25 upregulated and downregulated genes by Z scores.

(C) Pathway enrichment analysis by GO: molecular function.

(D) Validation of the expression of selected genes by qPCR; n = 3 mice.

(E and F) mRNA (E) and protein (F) expression of CX3CR1 in BMDMs with miR-130b/301b deletion (KO) and controls (WT) following engulfment of apoptotic thymocytes for 6 h. Bar graphs were shown as mean ± SD, n = 3 cell samples. ∗p < 0.05; ∗∗p < 0.01 (t test for D; ANOVA for E and F).

Role of CX3CR1 in migration of miR-130b/301b-deleted macrophages toward ACs

Peritoneal macrophages from miR-130b/301b KO and WT control mice were treated with or without the CX3CR1 antagonist, JMS-17-2. Migration toward ACs was assessed using the transwell cell chamber. As results shown in Figure 7, miR-130b/301b KO macrophages exhibited increased migration, whereas treatment with JMS-17-2 reduced peritoneal macrophage migration, suggesting a role of increased CX3CR1 in macrophages with miR-130b/301b deletion in enhancing macrophage migration toward ACs.

Migration of macrophages toward apoptotic cells

WT and miR-130b/301b KO mice were injected with 3.8% brewer’s thioglycolate followed by collection of peritoneal macrophages. Peritoneal macrophages treated with CX3CR1 antagonist or vehicle controls were loaded to the upper chamber with apoptotic thymocytes in the lower chamber. Representative images and cell count of migrated macrophages were shown (scale bars,100 μM). Bar graph was shown as mean ± SD. n = 3 cell samples in each group with three independent repeats. ∗p < 0.05 (ANOVA).

Global miR-130b/301b deletion attenuated high fat diet-induced adipose tissue inflammation

Given the implication of macrophage efferocytosis in obesity, we next determined the role of miR-130b/301b in the adipose tissue inflammation in high-fat diet-induced obesity. As results shown in Figure 8, high fat diet (HFD,45%, 12 weeks) increased the expression of proinflammatory factors, including Mcp-1 and Il-6 expression in iWAT of WT mice, which were significantly suppressed in miR-130b/301b KO mice. Expression of Tnfα was significantly lower in iWAT of miR-130b/301b KO mice with HFD (Figure 8A). Similar decreases in expression of Mcp-1, Il-6, and TNFα were observed in epididymal adipose tissue (eWAT) of KO mice following HFD (Figure 8C). Inefficient clearance of dead adipocytes contributes to the accumulation of crown-like structures (CLSs), which are clusters of macrophages surrounding dead adipocytes and a key feature of adipose tissue inflammation.²⁵^,²⁶^,²⁷ We next examined the CLSs in iWAT and eWAT tissue sections by co-staining of dead cells by TUNEL staining and macrophages using F4/80 antibody. As images shown in Figures 8B and 8D, dead cells were present in the CLSs of both iWAT and eWAT of mice following HFD. Consistent with the expression of inflammatory factors, the numbers of CLSs were lower in iWAT and eWAT of miR-130b/301b KO mice compared to WT mice (Figures 8B and 8D). Those results suggest a key role of miR-130b/301b in HFD-induced adipose tissue CLS accumulation and inflammation.

Decreased inflammation and CLSs in adipose tissue of mice with miR-130b/301b deletion

miR-130b/301b KO and WT male littermates were fed with HFD (45% kcal from fat, 20% kcal protein, and 35% kcal carbohydrate) at 5–6 weeks of age for 12 weeks.

(A and C) Total RNAs were extracted from inguinal fat (iWAT) and epidydimal fat (eWAT) tissues, and mRNA expression of *Mcp-1*, *Tnf-a*, and *Il-6* in iWAT (A) and eWAT (C) were measured by real-time PCR and normalized to TBP. n = 6–8 mice.

(B and D) Representative images of TUNEL and F4/80 co-staining and quantification of crown-like structures (CLSs) in iWAT (B) and eWAT (D) of WT or KO mice following high fat diet. CLSs were quantified in 3 random views (4x) from 3 mice in each group. Mean ± SD. #p < 0.1; ∗p < 0.05; ∗∗p < 0.01 (ANOVA for A and C; t test for B and D). Scale bars, 25 μm.

Discussion

Defective macrophage efferocytosis leads to prolonged inflammation, which plays a crucial role in the development of obesity and related metabolic disorders, including atherosclerosis and insulin resistance. We previously revealed dysregulation of miR-130b in adipocytes and ATMs,¹⁹ and an important role of the miR-130b/301b cluster in inhibiting beige adipogenesis and energy metabolism in obesity.²² Here, we further investigated the regulation and function of miR-130b/301b in macrophages. Utilizing in vitro and in vivo approaches, we found that miR-130b/301b expression was regulated by the inflammatory status of macrophages, and miR-130b/301b deletion promoted macrophage anti-inflammatory programming, mitochondrial respiration, and phagocytosis of ACs, attenuating adipose tissue inflammation in obesity.

The process of macrophage efferocytosis is associated with the production of anti-inflammatory factors, which are crucial for the resolution of inflammation and maintenance of tissue homeostasis.¹^,²^,³ We found that miR-130b/301b was downregulated following engulfment of ACs as well as by the anti-inflammatory stimulus IL-4. Deletion of miR-130b/301b in macrophages resulted in pro-inflammatory to pre-resolving anti-inflammatory polarization, and macrophages deficient in miR-130b/301b exhibited enhanced capability of phagocytosis of ACs. Moreover, clearance of ACs was enhanced in mice lacking miR-130b/301b. On the other hand, overexpression of miR-130b or miR-301b mimics in macrophages enhanced expression of inflammatory factors. Those results suggest that the downregulation of macrophage miR-130b/301b is an important event during the process of efferocytosis, which links to inflammation resolution. In the context of obesity, unresolved adipose tissue inflammation leads to insulin resistance and metabolic dysfunction. Here, global deletion of miR-130b/301b significantly decreased high-fat diet-induced adipose tissue inflammation in subcutaneous adipose tissue (iWAT) and eWAT of mice, which was associated with decreased CLs, supporting a link of enhanced macrophage efferocytosis by miR-130b/301b deletion to adipose tissue inflammation. As we published previously in the same cohort of mice study, miR-130b/301b deletion result in enhanced iWAT beigeing, decreased whole-body adiposity and glucose intolerance.²² The enhanced macrophage efferocytosis and anti-inflammatory programming due to the absence of miR-130b/301b can convey beneficial metabolic consequences in obesity and diabetes. In addition to obesity and diabetes, macrophage efferocytosis is critical for preventing cardiovascular disease by maintaining atherosclerotic plaque stability.²⁸ Given the effects on macrophage efferocytosis and inflammation resolution, miR-130b/301b can also be a promising therapeutic target for cardiovascular diseases.

Increased glucose metabolism, fatty acid oxidation, and mitochondrial oxidative phosphorylation play essential roles in IL-4-stimulated anti-inflammatory activation.²⁹^,³⁰ Consistent with the function of miR-130b/301b in adipose progenitor cells as reported previously,²² we showed that deletion of miR-130b/301b increased mitochondrial respiration in macrophages. Such an increase can consequently lead to anti-inflammatory polarization in macrophages. It has been demonstrated that metabolites and molecules derived from engulfed ACs fuel mitochondrial respiration and promote anti-inflammatory response following efferocytosis.¹²^,¹⁴ Reciprocally, mitochondrial energy metabolism and signaling are essential for efficient efferocytosis. Defective mitochondrial function in macrophages has been reported to cause impaired efferocytosis and suppress inflammation resolution.¹⁵ Therefore, miR-130b/301b deletion promotes macrophage efferocytosis and anti-inflammatory polarization likely via enhancing mitochondrial oxidative phosphorylation and function.

MicroRNAs play important gene-regulatory roles in cells by pairing with the mRNAs of protein-coding genes.¹⁶ We propose that miR-130b/301b affects efferocytosis and anti-inflammatory response via regulating the expression of PGC-1α and PPARγ in macrophages. miR-130b has been reported to directly bind to the 3′-UTR of both PGC-1α and PPARγ mRNAs.²¹^,²³^,³¹ PGC-1α is a key regulator of mitochondrial biogenesis and metabolism, which improves mitochondrial function and oxidative phosphorylation by upregulating genes involved in mitochondrial protein synthesis and energy production.³² Also, decreased PGC-1α exacerbates inflammation via promoting activation of the nuclear factor κB (NF-κB) pathway.³³ PPARγ, a ligand-activated transcription factor, induces macrophage anti-inflammatory polarization.³⁴^,³⁵ In the present study, overexpression of a mimic of miR-130b or miR-301b, but not negative control, significantly decreased protein abundance of both PGC-1α and PPARγ in macrophages. In contrast, macrophages deficient in miR-130b/301b exhibited increased expression of PGC-1α and PPARγ protein. Those results suggest an important role of miR-130b and miR-301b in regulating PGC-1α and PPARγ in macrophages, which can impact mitochondrial function, inflammatory response, and efferocytosis. Of note, miR-130b is reported to directly target AMP-activated protein kinase (AMPKα),²²^,³⁶ which is an important regulator of energy metabolism and macrophage efferocytosis.³⁷ However, in macrophages, protein abundance of AMPKα was not impacted by mimics of miR-130b or miR-301b, nor deletion of miR-130b/301b (data not shown), suggesting gene targeting of miR-130b/301b is cell-type specific.

A single miRNA or cluster can have the potent function as it can modulate the expression of multiple genes simultaneously, which link multiple cellular pathways.³⁸ Our unbiased RNA-seq data on bone marrow-derived macrophages showed that deletion of miR-130b/301b altered mRNA expression profiles in macrophages, which are largely involved in cell surface binding pathways, including adenyl nucleotides, immune cells, and growth factor binding. Notably, miR-130b/301b deletion in bone marrow-derived macrophages significantly increased expression of CX3CR1, a receptor of macrophages for the AC-released “find me” signal CX3CL1. CX3CR1 plays a crucial role in macrophage efferocytosis via triggering their migration toward ACs.³⁹ We found that deletion of miR-130b/301b enhanced macrophage migration toward ACs, which was attenuated by CX3CR1 antagonist. Hence, in addition to regulating mitochondrial metabolism, miR-130b/301b deletion can also facilitate the recruitment of bone marrow-derived macrophages to “find” ACs.

In summary, macrophage engulfment of ACs was associated with decreased miR-130b/301b expression, which consequently increased mitochondrial oxidative phosphorylation, anti-inflammatory polarization, and “finding” of ACs, thereby promoting efficient efferocytosis and inflammation resolution. miR-130b/301b can therefore be a promising therapeutic target against prolonged inflammation in chronic diseases, including obesity and related metabolic disorders.

Limitations of the study

We acknowledge the limitation of using global KO mice of miR-130b/301b for in vivo studies, which does not allow us to fully distinguish macrophage-intrinsic effects from potential systemic or adipocyte-derived influences. Future studies using macrophage-specific KO mice in HFD-induced obesity are warranted. Given the homology and functional redundancy between miR-130b and miR-301b, deleting both from the genome for our studies avoided the potential of one miRNA compensating for the other. However, this limits the capability to fully disentangle the functional contributions of miR-130b versus miR-301b, which remain to be studied using mice lacking each of them individually. Only male mice are included in all the experiments, which does not fully represent the entire population because of potential sex differences. Finally, our current findings on mice have not been validated on human macrophages. Translational relevance warrants future studies.

Resource availability

Lead contact

Further information and requests for resources should be directed to and will be fulfilled by the lead contact, Shaoning Jiang (shaoning.jiang@yale.edu).

Materials availability

This study did not generate new materials.

Data and code availability

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RNA-seq data are available via NCBI GEO database under accession codes Database: GSE312838 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE312838) by the time of publication. Original full western blots and additional supporting data are available in Mendeley Data (Mendeley Data: https://data.mendeley.com/datasets/7z7bgzsdtc/1). Accession codes are also available in the key resources table.
•
This paper does not report original code.
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Additional raw data will be made available from the lead contact upon reasonable request.

Acknowledgments

This study is supported by OCAST (principal investigator [PI], S.J.) and the National Institutes of Health (R01DK138986, PI, S.J.). Experiments using the XFe96 seahorse equipment were assisted by the Yale Diabetes Center Core Facility. Biorender access (graphical abstract) is supported by the Yale Liver Center. We acknowledge the support from the University of Oklahoma Pediatric Metabolic Research Program, Harold Hamm Diabetes Center, and Yale Department of Pathology.

Author contributions

All authors contribute to the conception, design, and interpretation of the data. S.J., L.Y., T.A., and Y.X. performed the experiments; G.C. did the RNA-seq analysis; S.J. and L.Y. wrote the manuscript. S.J. is the guarantor. All authors revised the manuscript and approved this version to be published.

Declaration of interests

The authors declare no conflict of interest.

STAR★Methods

Key resources table

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies

PPARγ	Cell Signaling Technology	Cat#2435; RRID: AB_2166051
PGC-1α	Cell Signaling Technology	Cat#4259; RRID: AB_2268435
β-actin	Cell Signaling Technology	Cat#5125; RRID: AB_1903890
CX3CR1	Abcam	Cat#Ab308613; RRID: AB_3271513
GAPDH	Proteintech	Cat#60004; RRID: AB_2920461
F4/80	Cell Signaling Technology	Cat#70076; RRID: AB_2799771
BV421-labeled anti-CD90.1	Biolegend	Cat#202529; RRID: AB_10899572
PE-CF594 F4/80	BD Horizon	Cat#565613; RRID: AB_2734770

Biological samples

Bone marrow derived macrophages	mice	In-house
Peritoneal macrophages	mice	In-house
Thymocytes	mice	In-house

Chemicals, peptides, and recombinant proteins

M-CSF	Biolegend	Cat#576406
Brewer thioglycolate medium	Sigma	Cat#B2551
CFSE	ThermoFisher Scientific	Cat#C34554
Dexamethasone	Sigma	D4902
JMS-17-2	MedChemExpress	Cat#JMS-17-2
miRNA mimics	Dharmacon Reagents	miRIDIAN Mimics
TNFα	PeproTech	Cat#315-01A-20UG
IL-1β	PeproTech	Cat#211-11B-10UG
IL-4	PeproTech	Cat# 214-14-20UG

Critical commercial assays

miRNeasy Advanced Mini Kit	QIAGEN	Cat#217604
TaqMan MicroRNA Reverse Transcription Kit	Life Technologies	Cat#4366596
Advanced miRNA cDNA Synthesis Kit	Life Technologies	Cat#A28007
High-Capacity cDNA Reverse Transcription Kit	ThermoFisher Scientific	Cat#4374967
TUNEL Apoptosis Kit	Novus Biologicals	Cat#NBP3-12093
Seahorse XF Mito Stress Test Kit	Agilent	Cat#103015-100
Power SYBR Green PCR Master Mix	Applied Biosystems	Cat# 4367659
Halt™ Protease and Phosphatase Inhibitor Cocktail	Thermo Scientific	Cat# PI78443

Deposited data

RNA-seq	NCBI GEO	GSE312838
Original WB data	Mendeley data	https://doi.org/10.17632/7z7bgzsdtc.1

Experimental models: Organisms/strains

Mouse: miR-130b/301b Global KO mice	Luo et al.²²	Diabetes. 2022 Nov 1;71(11):2360-2371. doi: https://doi.org/10.2337/db22-0205
C57BL/6J	The Jackson Laboratory	000664

Oligonucleotides

See Supplementary Table for primers	–	–

Software and algorithms

Flowjo software version 10	FlowJo LLC	https://www.flowjo.com/
GraphPad PRISM 5.0	GraphPad Software	https://www.graphpad.com/

Other

HFD diet	Research Diets	D12451i
Chow control diet	Research Diets	D12450Hi

Open in a new tab

Experimental model and study participant details

All mice experiments were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals and were approved by the Yale Institutional Animal Care and Use Committee (IACUC protocol #20570). The mice were kept on a 12-h light-dark cycle with free access to food and water. The miR-130b/301b global knockout (KO) mice was generated by crossing miR-130b/301b fl/fl mice (Jackson laboratory, no. 034655) with CMV-Cre mice (Jackson laboratory, no. 006054) as described in our previous study.²² Heterozygous miR-130b/301b KO male mice were bred with heterozygous female mice to produce homozygous KO mice and wild-type C57BL/6J controls for present experiments. Mice were euthanized by isoflurane followed by heart removal. Only male mice were used for in vitro cell isolation and in vivo experiments, including: (1) In vitro primary bone marrow and peritoneal macrophages isolation: Male miR-130b/301b KO mice and wild-type control mice under standard diet (6-9 weeks of age) were used for bone marrow or peritoneal macrophages isolation. The isolated cells were subjected to downstream assessments, including miR-130b/301b expression, macrophage polarization, in vitro efferocytosis, mitochondrial function, RNA-seq analysis, and trans-well migration (detailed methods were listed in following sessions in methods). (2) Male miR-130b/301b knockout and wild-type control mice with normal diet (12 weeks of age) were used for in vivo efferocytosis assessment (detailed method described below). (3) C57BL/6J male mice at 5-7 weeks of age with standard diet were used for tissue collection (iWAT, eWAT, BAT, quadriceps muscle, liver, and bone marrow) to assess tissue distribution of miR-130b following RNA extraction. (4) Same mice cohort of high fat-diet induced obesity model generated in our previous studies²² were used to study adipose tissue inflammation. Briefly, miR-130b/301b KO and control littermates (5–6 weeks of age) were fed HFD (D12451; Research Diets) with 45% energy from fat or a chow control diet (10% of kilocalories from fat, D12450Hi; Research Diets) for 12 weeks KO mice gained less body weight with no difference in food intake compared to WT controls. Fat depots (eWAT and iWAT) were dissected, weighed, and processed for histology studies (fixed in 4% PFA for 24 hours followed by paraffin embedding and sectioning) and RNA analysis (snap-frozen in dry ice, powdered in liquid nitrogen, and stored in -80 ^OC).

Method details

Isolation and culture of BMDM

Mice were sacrificed via isoflurane anesthesia followed by cervical dislocation. The lower limbs were dissected, and the surrounding muscles were removed. Bone marrow was collected and filtered through a 70-μm cell strainer. After red cell lysis, the bone marrow cells were cultured in DMEM/F12 medium supplemented with 10% FBS and M-CSF (10 ng/ml, Sigma). After 24 hours, non-adherent cells were collected and cultured in M-CSF (30 ng/ml) for an additional 3 to 5 days for macrophage differentiation. The differentiated macrophages were used for the indicated experiments.

Isolation and culture of peritoneal macrophages

Peritoneal macrophages were isolated as described previously.³⁷ Briefly, mice were intraperitoneally injected with 2 ml of 3.8% brewer’s thioglycolate (Sigma). Four days post-injection, the macrophages were harvested by peritoneal lavage and plated in 24-well plates (2 × 10⁵ cells/well). Non-adherent cells were removed by washing with culture medium after 1∼2 hours. Adherent macrophages were maintained in DMEM culture medium supplemented with 10% FBS and used for the indicated experiments.

In vitro efferocytosis assay

In vitro efferocytosis was performed as described previously with modification.³⁷ Briefly, thymocytes were collected from mice and labeled with CFSE (Life Technologies) for 30 min at 37°C according to the manufacturer’s protocol. The labeled thymocytes were treated with dexamethasone (1 μM) for 16 hours to induce apoptosis. The apoptotic thymocytes were washed thoroughly to remove dexamethasone and co-cultured with peritoneal macrophages (WT and miR-130b/301b KO) at a ratio of 5:1 for 2 hours. After co-culture, macrophages were stained with BV421-labeled anti-CD90.1 (marker for thymocytes) for 30 minutes on ice in the dark and analyzed by flow cytometer. Macrophages positive with CFSE, while negative with CD90.1 staining (CFSE⁺BV421^-), were gated as macrophages with engulfment of apoptotic cells. For efferocytosis of Hepa1-6 cells, apoptosis in Hepa1-6 cells with stable GFP expression was induced by heating at 50°C for 20 minutes. Peritoneal macrophages (WT and miR-130b/301b KO) were incubated with or without apoptotic for 1 hour, followed by flow cytometry analysis. F4/80⁺ cells with positive green fluorescence (FITC positive) were gated as macrophages with phagocytosis of apoptotic cells. All data analysis was performed using FlowJo software version 10 (FlowJo LLC, OR).

In vitro migration assay

Migration assay was performed using 3.0 μm pore size transwell chambers (Falcon). Briefly, 500 μl of peritoneal macrophage suspension (2 × 10⁵ cells/ml) was seeded into the upper chamber. After 2 hours, 2 × 10⁵ apoptotic thymocytes in 750 μl of medium were added to the lower chamber, with or without the CX3CR1 antagonist JMS-17-2 (20 nM; MedChemExpress LLC, NJ). Following 2 hours of incubation at 37°C in 5% CO2, the migrating macrophages on the lower surface were fixed with 4% paraformaldehyde for 30 minutes and stained with 1% crystal violet for 30 minutes. Cells in three random fields were imaged using an EVOS microscope (Thermo Fisher Scientific, MA).

In vivo efferocytosis assay

In vivo efferocytosis was performed as described previously with modification.⁴⁰ Male miR-130b/301b knockout and wild-type control mice were injected with dexamethasone (10 mg/kg) or vehicle (control, 300μl of PBS) intraperitoneally. Thymuses were dissected 16 hours after dexamethasone injection. To monitor the presence of uncleared apoptotic cells, thymuses were fixed in 4% paraformaldehyde for 24 hours and embedded in paraffin, and sectioned. TUNEL staining for apoptosis was performed using a kit (Novus Biologicals) according to the manufacturer’s protocol. In brief, the slides were deparaffinized and rehydrated. After washing with PBS, the slides were incubated with Proteinase K solution for 30 min. Then, the samples were incubated with 100 μL TUNEL Equilibration Buffer or enough to cover the sample for 5 min, followed by removing the equilibration buffer and adding 50 μL (or enough to cover the sample) of TUNEL reaction mix to each sample for 60 min at 37°C. The samples were observed under the fluorescence microscope.

Immunofluorescence analysis

The paraffin embedded eWAT and iWAT tissues were sectioned and deparaffined with xylene followed by rehydration with ethanol gradient. TUNEL staining was performed as described above in “In Vivo Efferocytosis Assay”. The sections were further incubated with F4/80 antibody (Cell signaling Technology, 70076) followed by incubation with Alexa-fluor 488-conjugated 2^nd antibody (Abcam). Fluorescence signals were assessed under the fluorescence microscope.

Transfection of miRNA mimics

The bone marrow derived macrophages were transfected with the miRNA mimics (Dharmacon Reagents) of miR-130b-3p (50 nmol/L) and miR-301b-3p (50 nmol/L) or equal amount of a scrambled negative control miRNA using Lipofectamine RNAiMAX Transfection Reagent (Life Technologies, Grand Island, NY) following the manufacturer’s protocol. Three days after transfection, the cells were activated with palmitate (0.5 mM) overnight to assess activation of inflammatory activation by real-time PCR as indicated in the figure legend. Efficiency of transfection (>95% cells transfected) was verified by fluorescent imaging.

Quantitative real-time PCR (qRT-PCR)

Total RNA was extracted using the miRNeasy Advanced Mini Kit (QIAGEN). The concentrations of isolated total RNA were determined by Nanodrop Spectrophotometers. For miRNA measurement, reverse transcription was performed using the TaqMan MicroRNA Reverse Transcription Kit or advanced miRNA cDNA Synthesis Kit according to the Applied Biosystems protocol (Life Technologies, Grand Island, NY). All qPCR reactions for miRNAs were performed using TaqMan Universal Master Mix II, no UNG (Life Technologies, Grand Island, NY). Relative expression of individual miRNA was calculated using the 2-ΔΔCT method, where Sno202 or U6 served as the endogenous control small non-coding RNA.

For measurement of mRNA expression, reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific), and real-time PCR (primer sequences in Table S1) was performed using SYBR Green Master Mix (ThermoFisher Scientific), and specificity was determined by performing a melting curve. Gene expression was normalized to TBP (TATA box binding protein) and calculated using the 2^−ΔΔCT method.

Western blot analysis

Cells were lysed in RIPA buffer in the presence of protease and phosphatase inhibitor cocktail (Pierce Biotechnology, Rockford, IL). Protein lysates were reduced in sample buffer (Sigma) and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The blots were incubated with antibodies specific for PPARγ (Cell Signaling Technology, #2435), PGC-1α (Cell Signaling Technology, #4259), β-actin (Cell Signaling Technology, #5125), CX3CR1 (Abcam, ab308613), and GAPDH (Proteintech, 60004), and detected by enhanced chemiluminescence (Pierce). The blots were quantified and analyzed by imaging densitometry with Image Lab Software (Bio-Rad, Hercules, CA).

Measurement of mitochondrial respiration

The oxygen consumption rates (OCRs) of WT and KO macrophages were measured using a Seahorse extracellular flux analyzer (XFe96; Agilent Seahorse). OCR was measured over time and after sequential injections of modulators (Seahorse XF cell mito stress test kit; Agilent), including oligomycin (Olig, ATP synthase inhibitor, 1 μM), phenylhydrazone (FCCP, uncoupler, 0.5 μM), and Antimycin A & Rotenone (mitochondrial respiration inhibitor, 1 μM). Indices including baseline OCR, proton leak, maximal respiration, and ATP production were determined.

RNA sequence analysis

BMDMs from WT and miR-130b/301b KO mice were collected, and total RNA was extracted. The quality of the RNA was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies), and total RNA was subjected to RNA sequencing analysis (the Yale Genome Center). Raw sequencing reads were processed using the nf-core/rnaseq pipeline (v3.18.0) [https://doi.org/10.5281/zenodo.1400710], for mapping and quantification, using the GRCm39 mouse reference genome and the Gencode vM36 transcriptome annotation on the Nextflow platform. The mapping was above 90% for all samples, resulting in more than 30M mapped pairs per sample, and PCA analysis revealed good clustering by group (WT and KO). Raw read counts were then processed using in-house scripts. A GLM within the edgeR framework was implemented to infer differentially expressed genes across conditions and used the trended dispersion to estimate the biological variance. Nominal p-values from differential expression analysis were FDR corrected, and an FDR cut-off of 0.05 was used for all the tests. ToppGene [doi: https://doi.org/10.1093/nar/gkp427] was used for functional enrichment analysis.

Quantification and statistical analysis

Data are shown as mean ± SD, unless otherwise specified. Statistical analyses were conducted using GraphPad PRISM 5.0 (GraphPad Software, San Diego, CA). Comparisons between two groups were conducted using analyses of unpaired t-tests, and comparisons among multiple groups were done by performing a two-way ANOVA analysis, followed by Turkey’s post hoc test for multiple comparisons. P<0.05 was considered statistically significant. ∗ P<0.05; ∗∗P<0.01; ∗∗∗P<0.001, and ∗∗∗∗P<0.0001. All the statistical details of experiments can be found in the figure legends.

Published: March 10, 2026

Footnotes

Supplemental information can be found online at https://doi.org/10.1016/j.isci.2026.115307.

Supplemental information

Document S1. Tables S1 and S2

mmc1.pdf^{(145.8KB, pdf)}

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Document S1. Tables S1 and S2

mmc1.pdf^{(145.8KB, pdf)}

Data Availability Statement

•
RNA-seq data are available via NCBI GEO database under accession codes Database: GSE312838 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE312838) by the time of publication. Original full western blots and additional supporting data are available in Mendeley Data (Mendeley Data: https://data.mendeley.com/datasets/7z7bgzsdtc/1). Accession codes are also available in the key resources table.
•
This paper does not report original code.
•
Additional raw data will be made available from the lead contact upon reasonable request.

[bib1] 1.Boada-Romero E., Martinez J., Heckmann B.L., Green D.R. The clearance of dead cells by efferocytosis. Nat. Rev. Mol. Cell Biol. 2020;21:398–414. doi: 10.1038/s41580-020-0232-1. [DOI] [PMC free article] [PubMed] [Google Scholar]

[bib2] 2.Sheng Y.R., Hu W.T., Chen S., Zhu X.Y. Efferocytosis by macrophages in physiological and pathological conditions: regulatory pathways and molecular mechanisms. Front. Immunol. 2024;15 doi: 10.3389/fimmu.2024.1275203. [DOI] [PMC free article] [PubMed] [Google Scholar]

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PERMALINK

Deletion of miR-130b/301b cluster promotes macrophage efferocytosis and resolution of adipose tissue inflammation

Lei Yang

Tuerdiguli Abuduyimiti

Yibo Xi

Gianfilippo Coppola

Chen Liu

Shaoning Jiang

Summary

Graphical abstract

Highlights

Introduction

Results

Decreased miR-130b expression following differentiation of BMDMs and in response to IL-4 anti-inflammatory stimulation

Figure 1.

Decreased miR-130b and miR-301b in BMDMs following phagocytosis of ACs

Deletion of miR-130b/301b cluster enhanced macrophage efferocytosis in vitro and in vivo

Figure 2.

miR-130b/301b regulates macrophage inflammatory polarization

Figure 3.

Deletion of miR-130b/301b enhanced mitochondrial respiration

Figure 4.

miR-130b significantly suppressed protein expression of PPARγ and PGC-1α

Figure 5.

Differential gene expression profiles and pathways in macrophages with miR-130b/301b deletion

Figure 6.

Role of CX3CR1 in migration of miR-130b/301b-deleted macrophages toward ACs

Figure 7.

Global miR-130b/301b deletion attenuated high fat diet-induced adipose tissue inflammation

Figure 8.

Discussion

Limitations of the study

Resource availability

Lead contact

Materials availability

Data and code availability

Acknowledgments

Author contributions

Declaration of interests

STAR★Methods

Key resources table

Experimental model and study participant details

Method details

Isolation and culture of BMDM

Isolation and culture of peritoneal macrophages

In vitro efferocytosis assay

In vitro migration assay

In vivo efferocytosis assay

Immunofluorescence analysis

Transfection of miRNA mimics

Quantitative real-time PCR (qRT-PCR)

Western blot analysis

Measurement of mitochondrial respiration

RNA sequence analysis

Quantification and statistical analysis

Footnotes

Supplemental information

References

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

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