Figure 2.

Single-cell RNA sequencing (scRNA-seq) of cardiac immune cells after pressure overload. A, Design of multiplexed scRNA-seq experiment using hashtag-oligos (HTO) barcoded antibodies to identify mouse of origin of each cell. B, Overview of identified barcodes used to select singlets for further analysis. C, Pie chart indicating the number of single cells sequenced for each condition (upper) and per mouse (lower). D, Uniform manifold approximation and projection (UMAP) projection of single cells clustered in 25 unique clusters with identification of immune cell identity. Sham and transverse aortic constriction (TAC)–derived cells are plotted separately to visualize abundance differences. E, Heatmap of top 5 identified genes that are specifically expressed within clusters using unsupervised clustering. F, Bar graph of relative abundance of each cluster of cells in sham vs TAC conditions (n=4, 6). Statistical significance between the sham and 1-wk TAC groups by cluster was determined by 2-tailed Mann-Whitney U tests. Data are presented as mean±SEM. Single-cell data is shown as scaled, variance-stabilized unique molecular identifiers (UMI) counts. DC indicates dendritic cell; NK, natural killer; and PMN, polymorphonuclear leukocytes. Statistical significance is summarized as ns, not significant and *P<0.05.