Fig. 5

The functional rescue of the Ccr5 ^−/− osteoclasts by active small GTPases. a Immunoblotting analyses of Vav3 and phosphorylated FAK were conducted 15 min after RANKL stimulation in wild-type and Ccr5 ^−/− BMCs. The immunoblotting data were replicated more than three times. b The levels of the active forms of Rac1, RhoA, and Cdc42 in osteoclastic differentiation were analyzed in cells from wild-type and Ccr5 ^−/− bones (mean ± SD, n = 5). c The adhesion ring formation of wild-type and Ccr5 ^−/− osteoclasts expressing the indicated constructs was examined in cells cultured on a glass-bottomed dish, and then analyzed by anti-vinculin immunofluorescence staining (shown in green, scale bars, 100 μm). Magnification ×20 (objective lens), n = 4. The vinculin intensity per cell perimeter was quantified and statistically compared (mean ± SD, n = 10). *P < 0.05 by Student’s t-test. d Resorption pit assays. After culturing, the osteoclasts were removed from the dentin slices and were subsequently stained with hematoxylin to visualize the resorption pits (scale bars, 100 μm, n = 5). The resorption pit areas (%) on images were scored and statistically compared. e The relative mRNA levels of Integrin-αV, Mmp3, and Mmp13 were measured by a real-time Q-PCR (mean ± SD, n = 4). *P < 0.05 by Student’s t-test