Fig. 1: Gonadal RANKL signaling and influence of OPG in mice.

a Triple immunofluorescence with RANKL (sc-9073, red), SOX9 (Sertoli cell marker, green), and VASA (germ cell marker, blue). b Double immunofluorescence with RANK (HPA0277728, green), VASA (red), and DAPI (blue), or c OPG (sc-21038, blue), SOX9 (green), and DAPI (red). d Triple immunofluorescence with RANKL (sc-9073, red), RANK (HPA0277728, green), OPG (sc-21038, blue), and DAPI (gray) (all IF in 16-week-old WT mice). White insert marks high magnification area. e, f Phenotypic effects of OPG (1 mg/kg, red) or vehicle (blue) injected twice weekly for 2 or 5 weeks on testicular and epididymal weight and total epididymal sperm count in 10-week-old mice, (e, left panel; p = 0.03, right panel; p = 0.04). Note, y-axes do not intersect x-axes at 0 (e, f). Scale bars correspond to 25 μm (a–d). Data are presented individually and as mean ± SEM, with n (vehicle/OPG) = 15/10 (left and middle panels, e) n (vehicle/OPG) = 9/10 (right panel, e), and n (vehicle/OPG) = 5/5 (all panels, f). Statistical test: two-sided Student’s t test (e, left and right panels) with *p < 0.05.