Fig. 1: Single-cell RNA-seq analysis of primate embryonic hearts.

a The tSNE analysis segregated a total of 1786 single cardiac cells, obtained from micro-dissected heart regions of primate embryos at 4 and 7 weeks of fetal age, into 13 clusters, including SHF/OFT progenitors (cluster #5). b Heatmap image depicting the representative differentially expressed genes in each of the 13 clusters in a. c Feature plots of the pan-cardiac and first heart filed (FHF)-related genes, as well as the SHF marker genes on the tSNE plots in a. d Violin plots of the same genes as in c in the segregated 13 clusters of the primate embryonic heart-derived single cells. e The rankings of the growth factor genes correlated with expression of the SHF-specific gene ISL1 (left) and the pan-cardiac/FHF-specific gene NKX2-5 (right) in single-cell RNA-seq data of primate embryonic hearts. The top 8 growth factors are highlighted in red shades, respectively. The corrected P-value for each gene was calculated by Guilt-by-Association and correlation analysis²⁶ (with Pearson correlation coefficient test). Source data are provided as a Source Data file. f Atlas of growth factor expression from multiple heart cell types in developing hearts (i.e., SHF, FHF, CM, pacemaker cell [PM], EC, SMC, and cardiac fibroblasts [CFB]), which was analyzed and defined by the Seurat and Guilt-by-Association and correlation analyses using single-cell RNA-seq data of primate embryonic hearts. In the Seurat program, each cluster (e.g., SHF: cluster #5, FHF: cluster #9, etc. [Supplementary Fig. 2a, b])-specific growth factors were identified, while in the latter, the growth factors correlated with each of the cell type-specific markers (i.e., SHF: ISL1, FHF: NKX2-5, CM: TNNT2; PM: SHOX2; EC: PECAM1, SMC: ACTG2, and CFB: DCN) were identified. The red, yellow, blue, and sky-blue colors indicate that each growth factor is ranked within the top 300 (red), top 301–600 (yellow), top 601–1200 (blue), or top 1201–2000 (sky-blue) genes correlated with each of the clusters and/or the cell type-specific markers. Thus, a red color in the chart indicates the strongest association between each of the growth factors and each of the heart cell types.