Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Jul 29;218(9):e20200667. doi: 10.1084/jem.20200667

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2021 DeBerge et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 3. — HIF-2⍺ suppresses anti-inflammatory macrophage mitochondrial metabolism. (A) IL-10 production by BMDMs or peritoneal elicited macrophages (PMac) from mice with myeloid-specific deletion of Hif1 (mHIF1^−/−), Hif2 (mHIF2^−/−), both Hif1 and Hif2 (mHIF1/2^−/−), or controls after efferocytosis of ACs. Nonengulfed cells were removed from adherent phagocytes. n = 3 sets of cells/group. Data are representative of more than three independent experiments. *, P < 0.05; ***, P < 0.001 by one-way ANOVA followed by Tukey’s test. (B) IL-10 production by BMDMs from mice with myeloid-specific overexpression of HIF-2⍺ (mHIF2^LSL) or controls after efferocytosis of ACs. n = 5 sets of cells/group. Data are representative of two independent experiments. *, P < 0.05 by two-tailed, unpaired t test. (C) Efferocytosis of calcein-labeled ACs (green) by MitoTracker-labeled BMDMs (red). Scale bar, 10 µm. n = 3 sets of cells/group. Data are representative of three independent experiments. ns, two-tailed, unpaired t test. (D) OCR with quantification of mitochondrial respiration by untreated (⍉) or efferocytic (AC) BMDMs. n = 6–8 sets of cells/group. Data are representative of three independent experiments. ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. AA/Ro, antimycin A/rotenone. (E) Expression of genes involved in fatty acid storage, synthesis, and oxidation in untreated mHIF2^−/− BMDMs or controls. n = 3 sets of cells/group. Data are representative of three independent experiments. *, P < 0.05; **, P < 0.01 by two-tailed, unpaired t test. (F) Expression of Hilpda in untreated BMDMs. n = 5 sets of cells/group. Data represent three independent experiments. *, P < 0.05; ***, P < 0.001 by one-way ANOVA followed by Tukey’s test. (G) IL-10 levels in cell culture medium after cocultivation of ACs and mHIF2^−/− BMDMs treated with control or Cpt1a siRNA. n = 3 sets of cells/group. Data are representative of two independent experiments. **, P < 0.01 by two-tailed, unpaired t test. (H) OCR with quantification of mitochondrial respiration by vehicle or TOFA-treated efferocytic BMDMs. n = 8 sets of cells/group. Data are representative of three independent experiments. *, P < 0.05 by two-tailed, unpaired t test. (I) IL-10 production by vehicle (Veh) or TOFA-treated efferocytic BMDMs. n = 5 sets of cells/group. Data are representative of three independent experiments. ***, P < 0.001 by two-tailed, unpaired t test. (J) BODIPY levels in BMDMs after cocultivation of BMDMs with BODIPY FL C₁₆-labeled ACs. n = 3 sets of cells/group. Data are representative of two independent experiments. ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. (K) BODIPY staining in BMDMs after cocultivation of BMDMs with BODIPY FL C₁₆-labeled ACs with quantification of lipid droplets (LD) per cell. Arrows indicate lipid droplets in BMDM. Scale bar, 10 µm. n = 4 sets of cells/group. Data are representative of three independent experiments. *, P < 0.05 by two-tailed, unpaired t test. (L) OCR with quantification of mitochondrial respiration by control or Hilpda siRNA-treated efferocytic BMDMs. n = 7–11 sets of cells/group. Data are representative of two independent experiments. **, P < 0.01; ***, P < 0.001 by two-tailed, unpaired t test. (M) IL-10 production by control or Hilpda siRNA-treated efferocytic BMDMs. n = 6 sets of cells/group. Data are representative of two independent experiments. ***, P < 0.001 by two-tailed, unpaired t test.