Figure 1.

Shn3^−/− mice have higher levels of CD31^hiEMCN^hi endothelium. (a, b) Representative confocal images (n = 4 total images per group) of 2-week-old Shn3^+/+ and Shn3^−/− male mouse femurs stained with CD31 (a) or Endomucin (EMCN) (b) (red) and 4′,6-diamidino-2-phenylindole (DAPI, blue). Growth plate and cortical bone are marked with a dashed line. Scale bars, 300μm (top) and 50μm (bottom). (c) Quantification of relative CD31-postive (top) and EMCN-positive vessel area (bottom) in the bone marrow (BM) cavity of the femur sections in 2-week-old Shn3^+/+ and Shn3^−/− male mice (n = 4 per group). (d) Representative images (n = 3 total images per group) of CD31 (green) and EMCN (red) dual immunostained femur sections from the femur in 2-week-old male mice. The growth plate is marked. Scale bars, 100μm. (e, f) Representative flow cytometry plots (e) with quantification (f) of CD31^hiEMCN^hi endothelial cells from the femurs of 2-week-old Shn3^+/+ and Shn3^−/− mice (n = 4 per group). Values represent mean ± s.e.m.; **P < 0.01, ***P < 0.001 by an unpaired two-tailed Student’s t-test in all panels.