Extended Data Fig. 9: at-RA modulates human MI-HSCs.
a, Left, Experimental design to characterize human sternal BM HSPCs of MI and Ctrl (control) donors after in vitro treatment with at-RA. Right, GSEA of published human HSPC signatures in human HSPCs upon in vitro culture with at-RA treatment vs. control (DMSO) from MI patients based on population RNA-seq. n = 2–4. b, GSEA of published human HSPC signatures in human HSPCs upon in vitro culture with at-RA treatment vs. control (DMSO) from MI patients based on population RNA-seq. n = 2–4. c, GO terms enrichment of upregulated DEGs (log2FC threshold = 0, P.adj < 0.1) in human MI-HSPCs with at-RA or 4-oxo-RA treatment vs. control (DMSO). d, ScHSPC division assay after 48 h in MI HSPCs after in vitro treatment with at-RA or control (DMSO). The percentage of cells is shown. Depicted p values correspond to the percentage of non-divided cells. Ordinary two-way ANOVA. n = 3. e, 1st and 2nd plating of human MI HSPC CFU assay after in vitro treatment with at-RA or control (DMSO). Unpaired t-test. n = 6. f, Volcano plots of DEGs between at-RA and DMSO (Ctrl) treatment in human BM HSPCs from healthy donors. DEGs that are common in previously published at-RA and DMSO (Ctrl) treatment in mouse BM HSCs (log2FC threshold = 0.5, P.adj < 0.1) are coloured in red (upregulated) or green (downregulated). 54 out of 313 upregulated human genes were conserved in mouse (17 %), while 78 out of 399 genes (20 %) were downregulated in both species. Important genes are annotated. g, GSEA of mouse RA direct target gene list in human HSPCs upon at-RA treatment vs. control (DMSO) from healthy donors based on population RNA-seq. h, Representative gating scheme for flow cytometric analysis of monocyte subtypes in PB of healthy donors. i, PCA of human PB monocytes from healthy donors upon control (DMSO), at-RA or 4-oxo-RA treatments based on the top 500 most variable genes in RNA-seq. n = 4. Data are presented as mean ± standard deviation. n indicates the number of biological replicates per condition or total number of human BM donors. For (d and e), three or more independent experiments were performed.
