Figure 5.

Osteoblast derived Slit3 controls osteogenesis and CD31^hiEMCN^hi endothelium via ROBO1. (a, b) Representative confocal images (n = 3 total images per group) of CD31 (red) and DAPI (blue) (a) or CD31 (green), EMCN (red) and DAPI (blue) (b) stained femur sections in 2 week-old OSX Cre and Slit3^osx male mice. Growth plate and cortical bone are marked with a dashed line. Scale bars, 100μm. (c) Representative μCT images of the trabecular bone in the distal femur metaphysis (left) and relative BV/TV analysis (right) of 3 week-old OSX Cre and Slit3^osx male mice. Scale bar, 1mm. n = 6 per group. (d) Representative μCT images of the femoral midshaft (left) and relative cortical bone thickness analysis (right) of 3 week-old OSX Cre and Slit3^osx male mice. Scale bar, 1mm. n = 6 per group. (e) Representative μCT images of the trabecular bone in the distal femur metaphysis (left) and relative BV/TV analysis in 12 week-old Robo1^+/+ (n = 4) and Robo1^−/− female mice (n = 5). Scale bars, 1mm. (f) Representative confocal images (n = 3 total images per group) of CD31 (green) and EMCN (red) with DAPI (blue) stained femur sections from 2 week-old Robo1^+/+ and Robo1^−/− mice. Growth plate is marked with a dashed line. Scale bars, 100μm. Values represent mean ± s.e.m.; *P < 0.05 and ***P < 0.001 by an unpaired two-tailed Student’s t-test in all panels.