Fig. 3. Effects of CTM, and gene silencing of Tgase-2 on the OPG expression in activated-MG-63 osteosarcoma cells. (A) CTM, dose-dependently suppressed the IL-1β-induced OPG expression of MG-63 cells. MG-63 cells were treated with CTM (0.1, 0.5, 1 mM) for 1 hr. Then, cells were treated with IL-1β for 24 h. Whole-cell lysates were prepared and the protein level was subjected to 4-12% SDS-PAGE, and expression of Tgase-2 and OPG were determined by Western blotting. (B) CTM, dose-dependently suppressed the TPA -induced OPG expression of MG-63 cells. MG-63 cells were treated with CTM (0.1, 0.5, 1 mM) for 1 hr. Then, cells were treated with TPA for 16 h. (C) Involvement of Tgase-2 in activated-MG-63 cells. MG-63 cells were transfected with siRNA of Tgase-2 (50 nM) for 48 hr. The cells were then treated with IL-1β (5 ng/ml) for 24 h. Whole-cell lysates were prepared and the protein level was subjected to 4-12% SDS-PAGE, and expression of Tgase-2 and OPG were determined by Western blotting.