Fig. 3.

Effects of nectandrin A on Id1 and Smad activation. (A) Id1 activation was measured by luciferase reporter assay using Id1-promoter-driven luciferase plasmid (Id1-Luc)-transfected C2C12 cells. Cells was cultured in a 6-well plate for 1 day and then incubated with DMEM containing 5% FBS in the presence or absence of rhBMP-2 with/without nectandrin A for 1 day. (B) The effect of nectandrin A on the activation of Smad was evaluated by Western blot analysis. Cells were cultured in a 6-well plate for 1 day and then incubated with DMEM containing 5% FBS in the presence or absence of rhBMP-2 (100 ng/ml) with/without nectandrin A for indicated time periods. (C) Cells were transfected with pCMV-Smad4-Flag with or without Id1-Luc reporter and then cells were incubated with DMEM containing 5% FBS and 100 ng/ml rhBMP-2 in the presence or absence of nectandrin A for 4 h. ^*p<0.05 compared to rhBMP-2 treated group.