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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. 6 — a Single-cell RNA sequencing of the epididymal white adipose tissue (EpiWAT) stromal vascular fraction (SVF) of C57BL/6 J mice following 6 or 12 weeks of high-fat feeding (Jaitin et al., 2019). Macrophages and monocytes were identified and expression of IRF5 and of GHITM were projected onto tSNE plots per cell type and duration of high-fat feeding. b Heatmap of mean expression values of IRF5, GHITM, ABCG1, SYCE2, FNIP2, ATF5 and LRRC27 over time and by cell type [monocytes (Mono) or macrophages (Mac)]. c Correlative analyses between IRF5 expression and expression of GHITM, ABCG1, SYCE2, FNIP2, ATF5 and LRRC27 at the single-cell level (Pearson’s correlation, Pearson r; two-tailed ***p < 0.0001 and **p = 0.004). Heatmap of IRF5 and GHITM expression, each line represents a single cell. d Heatmap of single-cell expression of IRF5, GHITM, ABCG1, SYCE2, FNIP2, ATF5 and LRRC27 over time and by cell type, each line represents a single cell. e Gene expression of GHITM in bone-marrow-derived macrophages (BMDMs) from mice with myeloid-restricted Cas9-GFP expression, treated with lipofection agent (Ctrl) or with a guide RNA (gRNA) targeting GHITM (gGHITM) and with or without Palm treatment for 2 h (n = 5 for Ctrl, n = 6 for other conditions, one-way ANOVA. ***p = 0.0003, left *p = 0.0423, right *p = 0.0167). Western blotting against GHITM in the same experimental design, quantification and blot in Fig. S8D, S8E (n = 2 per condition). f Oxygen consumption rate (OCR) from extracellular flux analysis in BMDMs with or without Palm treatment following transfection with gGHITM or with lipofection agent alone (Ctrl). Oligomycin (Oli), carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) and Rotenone/Antimycin A (Rot/AA) were administered (n = 5 for Ctrl, Ctrl+Palm and gGHITM; n = 8 for gGHITM + Palm). g Maximal respiration from extracellular flux analysis (n = 5 for Ctrl, Ctrl + Palm and gGHITM; n = 8 for gGHITM+Palm; one-way ANOVA, ****p = 0.000065 and **p = 0.0051). h Maximal respiration from extracellular flux analysis on Palm-treated BMDMs following transfection with a gRNA targeting IRF5 (gIRF5), double transfection with gGHITM and gIRF5 or with lipofection agent alone (Ctrl) (n = 3 per condition; one-way ANOVA, *p = 0.0428). Data presented as mean ± SEM. Source Data file provided.