Fig. 2

Cardiomyocyte transcriptome landscape upon CRISPRa Klf15. a Schematic showing the study design. CRISPRa mice with NT or Klf15 (gRNA) underwent either SHAM or TAC surgery to induce pathological cardiac hypertrophy. The hearts were harvested for scRNA-seq analysis (Dataset-3). b UMAP of cardiomyocyte subclusters (CM0-CM3) and stress score of single cardiomyocytes in the SHAM NT (gRNA), TAC NT or Klf15 (gRNA) hearts as defined by the expression of Nppa, Nppb, Acta, and Ankrd1. c Relative distribution of cardiomyocyte subclusters in the different conditions (SHAM NT (gRNA), TAC NT or Klf15 (gRNA) (CM0: pink; CM1: green; CM2: blue and CM3: violet). d UMAP of cardiomyocyte subclusters per condition. e Expression levels of Klf15 and EGFP reporter transcripts in the different cardiomyocyte subclusters (CM0-CM4). EGFP expression indicates AAV9 delivery. f Gene ontology (GO) of biological processes analysis of the different cardiomyocyte subclusters based on the significantly enriched genes (Log₂FC ≥ 0.25, P-value ≤ 0.05) in each subcluster. g Violin plot showing expression levels of KLF15 target Aldh2 and reprogramming marker Acta2, differentially regulated in the TAC NT and Klf15 (gRNA) groups. h Dotplot comparing KLF15-dependent network of genes contributing to Wnt and developmental signaling activation, hypertrophic pathways, Klf15 targets, and metabolic pathways, which are known to be controlled by KLF15, in different conditions (SHAM NT (gRNA), TAC NT and Klf15 (gRNA) groups) as well as in the different cardiomyocyte subsets (CM0-CM3). Different dot colors indicated a subcluster of CMs. Dot size indicates the proportion of cells expressing the gene, and color reflects the average z-scored expression level. i Representative immunofluorescence images of α-SMA and ACTN2 expression in SHAM NT (gRNA), TAC NT and Klf15 (gRNA) groups. α-SMA (red), ACTN2 (green), nuclei (DAPI, blue), scale bar = 20 μm. j Heatmap of proteomics data comparing the same groups depicting proteins representing main remodeling processes with corresponding gene ontology analysis (k). The Wilcoxon rank sum test was used for statistical significance in (e) and (g)