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. 2022 Aug 30;13:5089. doi: 10.1038/s41467-022-32813-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a CD14⁺ Monocytes from patients with type-2 diabetes (T2D; n = 5) were sorted based on expression of IRF5 for RNA-seq. Differential analyses were paired by patient and carried out on IRF5⁺ versus IRF5^- monocytes (n = 5, Wald test p-value < 0.05). b Gene ontology (GO) term enrichment from upregulated and downregulated genes in IRF5⁺ versus IRF5^- monocytes. c Expression of Ghitm in IRF5^- and IRF5⁺ monocytes (n = 5, *p = 0.039, two-tailed paired t-test). d Irf5 and Ghitm counts in white adipose tissue (WAT) macrophages and monocytes, from public dataset of scRNA-seq of the stromal vascular fraction (SVF) of patients that are lean or with obesity³² (two-tailed unpaired t-test, ****p < 0.0001). Heatmap of single-cell expression of Irf5 and Ghitm from monocytes (Mon) and macrophages (Mac), each line represents a single cell. e Proportion of Ghitm + (blue) and Ghitm− (red) cells in 10-cell bins by increasing Irf5 expression. f Correlation of Ghitm and Irf5 mean expression per bin (Pearson’s correlation Pearson R² = 0.28, two-tailed p < 0.0001). g Ghitm expression in CD14⁺ human visceral adipose tissue macrophages (vATMs). Samples were stratified based on expression of Irf5 into IRF5^Lo versus IRF5^Hi expressors (IRF5^Lo n = 7 and IRF5^Hi n = 6, two-tailed unpaired t-test, p = 0.13). h Correlation of IRF5 MFI, JC1-Green (mitochondrial mass, Mt Mass), JC1-Red (mΔΨ), and mΔΨ/mass in vATM from patients with obesity and in monocytes from patients with T2D (Mono) (n = 11 for monocytes, n = 9 for ATMs, Pearson’s correlation, Pearson r shown, p-values in Fig S9D). i Immunofluorescence staining of IRF5 (red), oxidative phosphorylation (OXPHOS) enzyme complexes (green) and CD14 (cyan) in human monocytes from patients with T2D. Nuclei stained visualised with DAPI (blue). Samples were separated based on IRF5 localisation, either nuclear (Nuc) or cytoplasmic (Cyt). Quantification of OXPHOS staining in Nuc and Cyt samples (n = 10 per condition, two-tailed unpaired t-test, *p = 0.0335). j University of California Santa Cruz (UCSC) genome browser^51–53 (http://genome.ucsc.edu) tracks at the GHITM locus. JASPAR2020⁵⁴ tracks visualise transcription factor binding sites for IRF5. BLUEPRINT^55,56 to visualise RNA expression and H3K27 acetylation marks in LPS-treated and -untreated human monocyte-derived macrophages (session link). Data presented as mean ± SEM. Source Data file provided.