Fig. 2: ATF3 as a key determinant for MerTK+ resident macrophage survival and/or proliferation in response to IR. | Nature Cardiovascular Research

Fig. 2: ATF3 as a key determinant for MerTK+ resident macrophage survival and/or proliferation in response to IR.

From: ATF3 coordinates the survival and proliferation of cardiac macrophages and protects against ischemia–reperfusion injury

Fig. 2

a, Flow cytometry gating strategy and quantification of Annexin V+ and EdU+ cells in MerTK+ macrophages in the heart of the sham operation and IR groups (n = 6 mice per group). b, Overlap between differentially expressed TFs and RCisTarget-predicted TFs (left). Violin plots of normalized scRNA expression profiles of the three TFs in two types of macrophages (right). c, Relative mRNA expression of Atf3, Jun and Fos in the heart tissues at different time points after IR (n = 6 mice per group). d, Relative mRNA expression of Atf3, Jun and Fos in MerTK+ and MerTK− macrophages sorted from the heart 6 h after IR (n = 6 mice per group). e, Flow cytometry analysis of Annexin V+ apoptotic cells in rGAS6-treated BMDMs transfected with the respective siRNAs (n = 6 biologically independent samples per group). f, Representative images (left) and quantification (right) of TUNEL-stained cells in rGAS6-treated BMDMs transfected with the respective siRNAs (n = 6 biologically independent samples per group). Scale bars, 50 μm. g, Flow cytometry analysis of EdU incorporation into rGAS6-treated BMDMs transfected with the respective siRNAs (n = 6 biologically independent samples per group). h, Representative images (left) and quantification (right) of Ki67+ cells in rGAS6-treated BMDMs transfected with the respective siRNAs (n = 6 biologically independent samples per group). Scale bars, 50 μm. i, Quantification of MerTK+, Trem2+, Lyve1+, MHCII+ and CCR2+ macrophages from the hearts of the two genotypes 6 h after sham operation and IR (n = 6 mice per group). j,k, Flow cytometry analysis of Annexin V+MerTK+ (j) and EdU+MerTK+ (k) macrophages in the heart of mice with two genotypes 6 h after IR (n = 6 mice per group). Statistical significance was evaluated via two-tailed, unpaired Student’s t-test (a, EdU; d, Fos, Jun; j and k), unpaired Mann–Whitney U-test (a, Annexin V; d, Atf3), Kruskal–Wallis test followed by Dunn’s multiple-comparison test (c), one-way ANOVA followed by Tukey’s multiple-comparison test (e–h) and two-way ANOVA followed by Tukey’s multiple-comparison test (i). All data are presented as mean ± s.e.m.

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