Fig. 1
From: Molecular determinants of nephron vascular specialization in the kidney
RNA sequencing analysis of kidney vascular endothelium throughout development. a Diagram denoting the workflow to sequence the bulk transcriptome of the vasculature throughout development. b Affinity propagation clustering of each stage. Edge lengths are proportional to Euclidean distances. Stages are color-coded according to the organ. c Diagram denoting the workflow to sequence the transcriptome of the vasculature at single-cell resolution. d Clustering of single-cell RNA expression according to a reduced dimensionality (t-SNE) for endothelial cells isolated from the kidneys of E17, P2, P7, and adult mice. VP, vascular progenitor; PTC, peritubular capillary; AA/LA, afferent arteriole/large arteries (pre-glomerular); AVR/V, ascending vasa recta/venous blood vessels; EA, efferent arteriole; GC, glomerular capillaries; DVR, descending vasa recta. e Heat map denoting genes enriched in each single-cell cluster. f Violin plots of normalized scRNA expression profiles of kidney endothelial cells. g Staining validation of Sox17 enriched in arteries in E15 kidneys. Scale bar 50 μm. h Staining validation of Gja5 in arteries except for the efferent arteriole in E15 kidneys. Scale bar 50 μm. i Flk1-eGFP reporter showing lower expression of VEGFR2 in arteries. Images were taken at the same exposure. Scale bar 50 μm. j Aqp1 staining in adult human kidney showing enrichment in afferent/pre-glomerular arterioles, and the descending vasa recta. Endothelial cells were marked with VE-cadherin (Cdh5) staining. Scale bar 100 μm. k FISH staining validation of Ehd3 showing enrichment in glomerular capillaries in E15 embryos. VR, Vasa recta. Scale bar 50 μm. l Staining validation of Plvap in peritubular capillaries, veins, and the ascending vasa recta in E15 kidneys. VR, vasa recta. Scale bar 50 μm. m Staining validations of Igfbp7 in the descending and ascending vasa recta in adult kidneys. Endothelial cells were stained with Endomucin (Emcn). PCT, proximal convoluted tubule. Scale bar left 100 μm, right 200 μm. n Staining validations of Igfbp5 in glomerular capillaries, peritubular capillaries, and the ascending vasa recta. CT, convoluted tubule. Scale bar 100 μm. o Illustration of known vascular subtypes which were identified through ddSEQ. Vascular subtypes not identified are grayed in the text below.
