Osteoclastogenesis Regulatory Proteins

Antibodies against the following proteins were used in this study: DOK3, C-Cbl, and Grb2 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Phospho-ERK (pThr²⁰²/pTry²⁰⁴), ERK (pan ERK), Phospho-p38 MAPK (Thr180/Tyr182), p38 MAPK (pan p38 MAPK), Phospho-Syk (Tyr525/526), Syk (panSyk), IκBα, cyclin D1 antibody were from Cell Signaling Technology (Danvers, MA, USA) ; Anti-phosphotyrosine monoclonal antibody G410 was purchased from EMD Millipore (Billerica, MA, USA). TRAP staining kit was purchased from Sigma (St. Louis, MO, USA). Phospho-Dap12 rabbit antibody was previously described (6 (link)). Murine M-CSF and RANKL were obtained from Peprotech (Rocky Hill, NJ, USA). The enhanced chemiluminescence (ECL) was purchased from Thermo Scientific (Rockford, IL, USA). Tissue culture media and fetal bovine serum (FBS) were obtained from Life Technologies (Grand Island, NY, USA). MTT (3-(4, 5-dimethylthiazole-2-yl)-2, 5-dipenyltetrazolium bromide) cell proliferation assay kit was obtained from ATCC (Manassas, VA, USA).