Extended Data Fig. 6: Endothelial-specific inactivation of Dach1.
From: Specialized post-arterial capillaries facilitate adult bone remodelling
a, t-SNE visualization of Dach1 gene expression in ECs of different organs. b, Confocal tile scan overview images of 12-week-old Dach1iΔEC and control retina stained for CD31 (red). Scale bars, 300μm. c, Quantitation of CD31+ vessel density in 12-week-old Dach1iΔEC retinas relative to control (n =4 mice per group). Mean ± SEM. P values, unpaired two-tailed t-test. d, High-resolution confocal images of Dach1iΔEC and control intestinal villi stained for DACH1 (green), VEGFR2 (red), and DAPI (blue). Scale bars, 50μm and insets 15μm. e, Quantitation of average VEGFR2+ vessel density, villus size, and average vascular area per villus in 12-week-old Dach1iΔEC and littermate control (n =4 mice per group). Mean ± SEM. P values, unpaired two-tailed t-test. f, Confocal tile scan overview images of 12-week-old female Dach1iΔEC and control femur stained for EMCN (red) and OSTERIX (green, OSX). White dashed lines and yellow arrowheads indicate reduction of region containing trabecular bone (TB) in Dach1iΔEC mutants. Scale bars, 500μm. g. High-magnification images showing EMCN (red) and ATP6V1B1/B2 (green) immunostained female Dach1iΔEC and control femur. Growth plate (GP) is indicated. Scale bars, 100μm. h. Quantitation of OSX+ cells and ATPV6B1B2 area in control and Dach1iΔEC mutant (n=3 and 4 mice per group). Mean ± SEM. P values, unpaired two-tailed t-test. i. Quantitation shows connectivity density and cortical thickness (per mm), (n = 3-4 female mice per group and n=3 male mice per group). Mean ± SEM. P values, plotted using unpaired two-tailed t-test. j. DACH1 (green) co-stained with VE-CADHERIN (red) in siCONTROL and siDACH1 cultured HUVECs. k. Downregulation of arterial marker genes in siDACH1 HUVECs relative to siCONTROL. n=6 (3 independent experiments) for each group. Mean ± SEM. P values, unpaired two-tailed t-test with Welch’s correction.
