Fig. 4
From: Molecular determinants of nephron vascular specialization in the kidney
Tbx3 is necessary for glomerular morphogenesis and specification. a Heat map denoting SCENIC results. 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. b Violin plot and tSNE showing normalized Tbx3 expression. c Immunofluorescent staining of Tbx3 and VE-cadherin (Cdh5) in adult human glomerular endothelial cells. EC, endothelial cell; Non-EC, non-endothelial cell. Scale bar 50 μm, inset scale bar 10 μm. d Masson’s trichrome staining in control and Tbx3ΔEC. Yellow arrow show microaneurysms. Scale bar 50 μm. e Pie chart for percent of glomeruli possessing microaneurysms, hypoplasia, or fibrosis. f Quantification glomerular capillary area. n = 3 mice, 5 frames/kidney. ***p < 0.0001 unpaired Student t-test. Error bars, standard error of the mean. g–l Urine analysis (control n = 15, Tbx3ΔEC n = 19). g Urea nitrogen, h creatinine (CREA), i micro total protein (MTP), j sodium (Na), k chloride (CL), l potassium (K). Normalized to volume. * p < 0.05, **p < 0.01, ***p < 0.001 unpaired Student t-test. Error bars, standard error of the mean. m Transmission electron microscopy (TEM) of control and Tbx3ΔEC. Red dotted line outlines glomerular capillary lumens. P, podocyte; GBM, glomerular basement membrane; EC, endothelial cell; RBC, red blood cell; L, leukocyte. Scale bar 2 μm, inset scale bar 500 nm. n Systolic blood pressure (n = 6 mice/group). *p < 0.05 unpaired Student t-test. Center line = median. Bounds of boxes: the first to third quartile. Whiskers highlight quartile from minimum or maximum. Error bars, standard error of the mean. o–q qPCR of Renin (Ren), Angiotensinogen (Agt), and Angiotensin-converting enzyme (Ace) transcripts (kidneys, liver, and lung, respectively). n = 4 mice each group. *p < 0.05 unpaired Student t-test. Error bars, standard error of the mean. r Hypergeometric test p-values (−1*log10) for overlap between mouse glomerular genes (scRNA sequencing) and genes down (blue) or up (green) regulated upon overexpression of the indicated transcription factors. EC, endothelial cell. s Euler plot illustrating overlap between glomerulus genes downregulated with overexpression with transcription factor. t Hypergeometric test p-values (−1*log10) for overlap between human glomerular-specific genes (human glomerular endothelial cells) and genes downregulated upon overexpression of the indicated transcription factors (ALL = all 4 TF’s). EC, endothelial cell. Red dotted line denotes p-value >0.05. u Pathway enrichment analysis after over expression of Tbx3, Gata5, Prdm1, and Pbx1.
