Fig. 1: Single-cell transcriptomic profiling and trajectory analysis of cardiac macrophages in a murine model of NICM.

a Dynamics of cardiac macrophages (live CD45+ CD64+ CD11b+ F4/80+ Ly6G−) from B6J mice treated with Ang-II (500 ng/kg/min). Cardiac macrophages are shown as a percentage with respect to total number of cardiac cells. Three biological replicates were used at each time point. P-values calculated by Mann–Whitney U test, vs percentage at day 0. *P < 0.05, **P < 0.01. b Cardiac CD45+ cells sorted from left ventricle (LV) from B6J mice treated with Ang-II (500 ng/kg/min, 7 days) and saline (controls) were pooled for scRNA-seq analysis on 5497 cells. Global uniform manifold approximation and projection (UMAP) dimension reduction analysis identified six major cell types. DC dendritic cells, NK cells natural killer cells. c Macrophages were re-clustered using Seurat’s Louvain clustering. d Heatmap of the scaled gene expression of top 10 markers for each macrophage cluster from Fig. 1c, based on average log₂ fold change (FC). ISGs Interferon (IFN)-stimulated genes, AP-1 Activator protein 1. e Heatmap of enriched pathways identified in each cardiac macrophage cluster. The top five terms are shown, and the significance of enrichment is represented by log₂(−log₁₀ (adjusted p value)). f Bar plot showing the relative proportions of macrophage clusters from c in B6J control (saline) and B6J mice treated with Ang-II (500 ng/kg/min, 7 days). A two-proportion z test was used to assess statistical significance; ***P < 0.001 Ang-II-treated vs saline-treated control mice. g Monocle-derived pseudotime cell trajectory of each cardiac macrophage cluster from c. h UMAP of cardiac macrophages describing developmental trajectories superimposed (upper) on clusters from C0 to C4 (lower). The main trajectories are generated by Slingshot, which are calculated as the average pseudotime over all cells.