Fig. 6: Expression of gene markers for macrophage and lymphatic endothelial cell populations.

Schematic of the generation of scRNA-seq datasets of control and injured P1 versus P7 CD1 mice. Hearts were harvested at 1 day after MI (P1MI1dpi and P7MI1dpi) or 7 days after MI (P1MI7dpi and P7MI7dpi). For the intact conditions, the samples were collected at either P1 or P7 (Intact P1 and Intact P7). The samples were FACS sorted using 7-AAD to isolate live cells, and libraries were prepared for sequencing using the 10x Genomics platform (a). For each timepoint, one library was generated using pooled tissues dissected from three individual animals to control for differences among individual animals, surgery and tissue dissociation variations. UMAP plot showing the different major clusters, ‘heartsClu2’, in two dimensions (b). To validate the clustering, known lymphatic-associated and macrophage-associated genes were examined using the integrated scRNA-seq dataset (c). Unbiased Gene Ontology analysis identified pathways upregulated in macrophages after injury at P7 compared to P1 (d) and genes potentially driving these pathways (e). Significant differences between Gene Ontology term enrichment were calculated using Fisher’s exact test. f, Heatmap of changes in lymphatic endothelial cell gene expression across conditions. Panel a created with BioRender.com. Clusters: 0-EC1 (endothelial cells 1), 1-FB1 (fibroblasts 1), 2-Mac, 3-FB2 (fibroblasts 2), 4-EC2 (endothelial cells 2), 5-EC3 (endothelial cells 3), 6-FB3 (fibroblasts 3), 7-Granulocytes, 8-SMC (smooth muscle cells), 9-Pericytes, 10-FB4 (fibroblasts 4), 11-EC4 (endothelial cells 4), 12-TC (T cells), 13-FB5 (fibroblasts 5), 14-EC5 (endothelial cells 5), 15-EC6, 16-BC (B cells), 17-CM (cardiomyocytes), 18-unassigned, 19-Epi (epicardium), 20-FB6 (fibroblasts 6), 21-Glial (glial cells), 22-unassigned, 23-Mo (monocytes).