Fig. 2: EC-specific deletion of ZEB1 reduces bone angiogenesis.
From: Endothelial ZEB1 promotes angiogenesis-dependent bone formation and reverses osteoporosis
a, b Representative confocal images of CD31/EMCN immunostaining in tibia of juvenile 3-week-old Zeb1ΔEC mice (a; n = 5) and Zeb1iΔEC mice (b; n = 5, each), and their corresponding littermate controls (n = 5, each). Magnified images of type H and type L vessels are shown in middle and right panels (a), respectively. Scale bar, 100 μm. Arrowhead denotes interface between type H and L vessels. c Quantification of type H and type L vessel densities in tibia as described in a and b (n = 5 independent experiments). d FACS plots of CD31 and EMCN double-stained single-cell suspensions from 3-week-old Zeb1WT and Zeb1ΔEC tibia (n = 5, each). e Quantification of type H and type L bone ECs in LIN− (i.e., DAPI−Ter119−CD45−) cells as shown in d (n = 5 independent experiments). f Confocal images of 3-week-old Zeb1WT and Zeb1ΔEC tibia (n = 5, each) immunostained with VEGFA and EMCN. Scale bar, 100 μm. Arrowhead marks VEGFA positively staining in endothelial column/arches, surrounding perivascular cells, and mature/hypertrophic chondrocytes. g Quantification of VEGFA staining levels in tibia as shown in f (n = 5 independent experiments). h RT-qPCR analysis of Vegfa transcript in FACS-sorted bone ECs of 3-week-old Zeb1WT and Zeb1ΔEC mice (n = 3 independent experiments). i Representative angiographic images of vascular organization of 3-week-old Zeb1WT and Zeb1ΔEC tibia (n = 5, each). Dashed boxed sections denote vessels near metaphysis. Scale bar, 0.5 mm. j Confocal images of EMCN immunostained distal columns and arches next to gp in 3-week-old Zeb1WT and Zeb1ΔEC tibia (n = 5, each). Arrowhead denotes filopodia extension. Magnified areas of dashed boxed sections are shown in bottom panels. Scale bar, 20 μm. k Quantification of filopodia numbers in tibia as shown in j (n = 5 independent experiments). All data are represented as mean ± SD. **P < 0.01; NS not significant. Differences are tested using unpaired two-tailed Student’s t-test. The source data are provided as a Source Data file.
