Fig. 1: Ephrin-B2 and EphB4 regulate EC segregation and artery formation.
From: Eph-ephrin signaling couples endothelial cell sorting and arterial specification
a Homotypic or heterotypic co-culture of HUVECs and HUAECs, as indicated. Final snapshots of CellTracker-labeled cells imaged for 48 h after removal of Ibidi insert. Quantitation graph for relative border length (n = 3 experiments). b RT-qPCR for EPHB4 and EFNB2 in HUVECs and HUAECs (n = 3 experiments). c, d EphB4 restrains while ephrin-B2 promotes artery growth. High-magnification confocal images of IB4 and GFP stained P6 retinal vasculature (n = 6 control and Ephb4iΔTC mice (c) and n = 6 control and 4 Efnb2iΔTC mice (d)). e, f Quantitation of the GFP+ area (Esm1-derived progeny) in capillary plexus, front and artery per total GFP positive area (n = 6 control and Ephb4iΔTC mice (e) and n = 6 control and 4 Efnb2iΔTC mice (f )). g, h Arteries in Ephb4iΔTC mice are longer and have more branches, while both parameters are reduced in Efnb2iΔTC mice. Graphs show number of arterial branches and artery extension relative to vascular plexus extension (n = 9 control and 8 Ephb4iΔTC mice (g) and n = 6 control and 4 Efnb2iΔTC mice (h)). i Representative images of Ephb4iΔTC retinas with arterial alterations compared to control. Graph shows quantitation of AV retinal crossings, overextended arterial side branches and normal vascular morphology (n = 17 control and 17 Ephb4iΔTC). j Scheme depicting that ephrin-B2-EphB4 interaction and correct levels are required for proper artery formation. P values were calculated by one-way ANOVA (a), two-tailed unpaired t test (b–h) and Chi-square test (i). In vivo experiments were performed with tamoxifen injections from P1-P3 with analysis at P6 (c, d, g–i). Data are presented as mean values ± SEM. Source data are provided as a Source Data file.
