Extended Data Fig. 4: at-RA treatment modulates HSC activity post-MI but leads to accumulation of cardiac myeloid cells.
a, Detailed experimental design to characterize HSC response after in vivo at-RA treatment following MI. Readouts are shown at precise time points throughout acute, reparative and chronic phases of MI. b, GSEA of mouse RA metabolism signature in pairwise comparisons vehicle vs. sham and at-RA vs. vehicle HSCs at day 2 post-MI population RNA-seq. c, GSEA of Reactome pathways in at-RA vs. vehicle HSCs upon MI. d, Representative gating scheme for flow cytometric cell cycle analysis of HSCs. e, 1st, 2nd and 3rd plating of HSC CFU assay comparing sham, MI+vehicle and MI+at-RA condition. Two-tailed unpaired t-test. n = 5–11. f, Representative gating scheme for flow cytometric analysis of cardiac leukocytes during EH. g, Left, Flow cytometry-based analysis of HSPC cell frequencies during emergency hematopoiesis in the acute phase at day 2 post MI in BM. Center, Flow cytometry-based analysis of myeloid cell and macrophage cell frequencies in the myocardium during emergency hematopoiesis in the acute phase at day 2 post MI in BM. Right, Cardiac macrophage frequency normalized to mg of infarct tissue is depicted. Ordinary one-way ANOVA. B(M) n = 8–18. h, Representative images of myocardium CD11bpos immunohistochemistry (IHC) stainings of vehicle and at-RA condition in the chronic phase at day 28 post MI. RZ, Remote zone; BZ, Border zone; IZ, Infarct zone. Infiltrated CD11bpos are pointed with an arrow. i, Representative gating scheme for flow cytometric isolation of cardiac cell populations isolated from mice after in vivo at-RA or vehicle (DMSO) treatment. In (b-g) cells were isolated in the acute phase at day 2-3 after MI. Data are presented as mean ± standard deviation. n indicates the number of biological replicates per condition. For (e,g), three or more independent experiments were performed.
