Distinct profile of E10.5 and E11.5 phenotypic HE. (A) Schematic representation of cell populations at different stages of EHT in dorsal aorta (DA) alongside corresponding phenotypic markers used for FACS single-cell sorting. (B) Uniform Manifold Approximation and Projection (UMAP) plots showing all the cells isolated from the E10.5 and E11.5 DA. FACS populations (left) and in silico clustering (right). (C) Top: Distribution of FACS-isolated populations in each cluster with background shaded according to the embryonic day of the samples. Bottom: Heat map illustrating the hematopoietic and endothelial gene expression profile of each cluster. Genes were clustered based on Pearson correlation, whereas the cells were grouped according to the clustering results. (D) Gene ontology (GO) analysis of genes upregulated in E11.5 FACS-HE^Gfi1-het compared with E10.5 FACS-HE^Gfi1-het. (E) Violin plot combined with box plot showing the normalized expression (log counts) of venous (Aplnr), arterial (Fbln5), and arterial fate promoting (Notch1, Jag1, Sox7, Sox17) genes in indicated populations. The shaded background behind the plot indicates the embryonic day origin of the cells. Medians are shown as a solid black line; gray boxes indicate the upper (75%) and lower (25%) percentile. *P < .05, **P < .01, ***P < .001 (2-tailed Student t test). NS, not significant.