Figure 3.

CD8⁺T cell depletion promotes early changes in cardioprotective metabolic gene expression after TAC. (A) UMAP plots of cardiomyocytes from sham, WT, CD3ϵ-/-, and CD8⁺T cell depleted mice at 2 weeks after TAC. Cells (dots) are colored for each group. scRNAseq analysis was performed as described in the Materials and Method section. (B) Box plots of representative expression of each gene in cardiomyocytes (Myh7; myocardial hypertrophy, Igf1r; growth factor receptor, Csf1; macrophage induction). Statistical differences were analyzed with a Wilcoxon rank sum test. (C) Mean decrease in accuracy (in order of decreasing accuracy from top to bottom) of each module eigengene (ME) as assigned by the random forest classifier. Random Forest algorithm was performed on 45 modules identified by weighted gene co-expression network analysis (WGCNA) restricted to 11,979 genes expressed at a fragments per kilobase of exon per million reads mapped (FPKM) value of ≥5 in at least 5 cardiomyocytes from a total of 21,859 genes. ME; the first principal component of the expression profile of genes belonging to the module. (D) UMAP plots colored by the expression of each module. (E, F) List of genes constituting M6 (E) and M12 (F). (G) Single-cardiomyocyte trajectory on the UMAP plot reconstructed by Slingshot. The lower right part, where cardiomyocytes from sham mice were enriched, was defined as the starting point. (H) Heatmap showing expression of M6/12 genes and cardioprotective cytokine receptor genes in pseudo-time order.