Osteoclast Differentiation Protocol

Acacetin and linarin were purchased from Sigma-Aldrich Chemical (St. Louis, MO, USA), as were all other reagents, unless specifically stated elsewhere. Antibodies of mouse CAII and ClC-7 were provided by Abcam Biochemicals (Cambridge, UK). Antibodies of mouse V-ATPase, mouse MMP-9, mouse cathepsin K, mouse integrin αv, mouse osteocalcin, mouse osteopontin, mouse TNSALP, and mouse collagen type I were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse integrin β3 antibody was purchased from Cell Signaling Technology (Beverly, MA, USA). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG, donkey anti-goat IgG, and goat anti-mouse IgG were supplied by Jackson ImmunoResearch Laboratory (West Grove, PA, USA). Mouse monoclonal β-actin antibody was purchased from Sigma-Aldrich Chemical (St. Louis, MO, USA).