Fig. 4: c-Src K152R mutation affects osteoclastic bone resorption.

a Model of the interaction between the SH2 domain of c-Src and RACK1. Key interacting residues were rendered as a space-filling structure. b First, 293T cells were transfected with the indicated expression vectors. RACK1 was immunoprecipitated from cell lysates using anti-HA. The precipitated complexes and cell lysates were analyzed using western blotting. The ratios of c-Src to HA-RACK1 and p-c-Src to c-Src were quantified in each of three independent experiments. *P < 0.01. c BMMs transduced with pMX-puro empty vector (EV), pMX-puro-WT-c-Src (Src WT), or pMX-puro-K152R-c-Src (Src K152R) were cultured for 3 days with 30 ng/mL M-CSF and 100 ng/mL RANKL to generate mature osteoclasts. Mature osteoclasts were seeded on bone slices and cultured for 3 days. The cells were then removed, and the bone slices were stained. Scale bar, 50 μm. Right: images of the stained sections were used to calculate the resorption pit areas. Data are presented as the mean ± SD of three independent experiments. *P < 0.01, **P < 0.05