Fig. 3. Inhibitory effects of mCHL2 on BMP-dependent differentiation of C2C12 cells in vitro. Dose-dependent inhibition by CHL2, chordin or noggin of BMP-dependent alkaline phosphatase (AP) induction in C2C12 cells. (A) Cells were cultured in differentiation medium for 2 days with various concentrations of BMP4 (plus sign), BMP6 (circle), or BMP7 (triangle). (B-D) Cells were also differentiated with a constant concentration of BMP4 (10 nM; B), BMP6 (15 nM; C) or BMP7 (19 nM; D) and various concentrations of mCHL2-FLAG (plus sign), noggin-Fc (triangle), or mCHD-His (circle) for 2 days. Mean specific activity of AP (triplicate assays) is shown; vertical bars denote s.d. Data are representative of three independent experiments.