Fig. 3: ATF3 controls MerTK+ macrophage fate via inhibiting type I IFN.
a, GSEA comparing rGAS6-stimulated BMDMs from ATF3-CKO and WT mice (n = 3 mice). NES, normalized enrichment score. b, Heatmap of DEGs in type I IFN signals. c,d, Flow cytometry analysis of Annexin V+ and EdU+ cells in sorted MerTK+ macrophages treated with rIFNα (c) or rIFNβ (d; n = 6 biologically independent samples). e, Pie chart of the percentage of peaks within each category bound by ATF3. f, Genome browser view of ChIP–seq tracks for ATF3 at the Ifih1, Ifnb1 and Apaf1 loci in BMDMs. g, ChIP–qPCR analysis of ATF3 binding to the Ifih1, Ifnb1 and Apaf1 promoter regions in BMDMs (n = 6 biologically independent samples). The signal was relative to the percentage input. h, Relative mRNA expression of Ifih1, Ifnb1 and Apaf1 in rGAS6-induced BMDMs infected with LV-Con or LV-ATF3 (n = 6 biologically independent samples). i, Relative mRNA expression of Ifih1, Ifnb1 and Apaf1 in rGAS6-stimulated BMDMs from ATF3-CKO and WT mice (n = 6 biologically independent samples). j, Relative mRNA expression of Ifih1, Ifnb1 and Apaf1 in MerTK+ and MerTK− macrophages sorted from the heart of ATF3-CKO and WT mice 6 h after IR (n = 6 mice). k, Outline of IFNAR NAbs (20 mg kg−1 d−1) or IgG treatment in ATF3-CKO mice subjected to IR surgery. l, Relative mRNA expression of Ifit1, Ifit2 and Ifit3 in the hearts of the two groups (n = 6 mice). m, Representative IF images and quantification of MerTK+ macrophages in the hearts of the two groups (n = 6 mice). Scale bars, 20 μm. n, Representative flow cytometry plots and quantification of MerTK+ macrophages in the hearts of the two groups (n = 6 mice). o,p, Flow cytometry analysis of EdU+ (o) and Annexin V+ (p) cells in MerTK+ macrophages in the hearts of the two groups (n = 6 mice). Statistical significance was evaluated using two-tailed, unpaired Student’s t-test (g, Apaf1; h, Ifih1, Ifnb1; i and l, Ifit2; and m–p), unpaired Mann–Whitney U-test (g, Ifih1, Ifnb1; h, Apaf1; l, Ifit1, Ifit3), one-way ANOVA followed by Tukey’s multiple-comparison test (d, EdU), Kruskal–Wallis test followed by Dunn’s multiple-comparison test (d, Annexin V) and two-way ANOVA followed by Tukey’s multiple-comparison test (j). All data are presented as mean ± s.e.m. Ctl, Control.
