Fig. 4.

Increased osteoblast maturation in rank^−/− mice. a Light micrographs of TRAP staining performed on trabecular bone sections from distal femora. Scale bar = 100 μm. b Formation of TRAP-positive cells from bone mononuclear cells isolated from rank^−/− and rank^+/+ mice. Scale bar = 100 μm. c Quantification of mature osteoclasts. d Serum TRAcp5b levels of 8-week-female adult rank^+/+ and rank^−/− mice. e Osteocalcin immunohistological staining of the metaphyseal trabecular bone at distal femora. Scale bar = 100 μm. f Serum OCN levels of 8-week-female adult rank^+/+ and rank^−/− mice. g–h Immunohistochemical analysis of runx2 in the metaphyseal trabecular bone at distal femora. Scale bar = 20 μm. i–j Immunohistochemical analysis of osterix in the metaphyseal trabecular bone at distal femora. Scale bar = 20 μm. All data are mean ± s.e.m. n = 8. *P < 0.05, **P < 0.01 by one-way ANOVA followed by t-test