Fig. 3

Prenatal dexamethasone exposure (PDE) induces loss of Nestin⁺ cells and type H vessels in the long bone of female mouse offspring. a Representative images of immunofluorescence staining for Nestin (green) and b quantitative analysis of Nestin⁺ cells in femoral metaphysis from 2-, 4-, and 6-week-old female offspring. 2W, 4W, and 6W represent 2-, 4-, and 6-week-old mice, respectively. DAPI stains nuclei blue. Scale bar, 30 μm. c Representative images of double-immunofluorescence staining for CD31 (green) and Endomucin (Emcn, red) in femoral metaphysis from 2-, 4-, and 6-week-old female offspring. DAPI stains nuclei blue. Scale bar, 40 μm. d Quantification of the relative fluorescence area of CD31⁺Emcn⁺ cells per tissue area in femoral metaphysis (CD31⁺Emcn⁺ area per mm²). Data are represented as mean ± S.E.M. *P < 0.05 versus control (n = 5 per group, Student’s t test)