BMP activity in ovine serum.
A, C3H-B12 cells were exposed overnight to different dilutions of ovine serum before assaying for luciferase activity. Values are the mean ± S.E. from one representative experiment with duplicate determinations. These experiments were performed three times with similar results. C3H-B12 cells were exposed overnight to ovine serum ½-diluted supplemented with increasing doses of noggin from 0 to 4 × 10−10
m, 2.5 × 10−6
m dorsomorphin (DM) or 10−8
m follistatin (B) or pituitary CM (CM basal 48 h) diluted ½ (D) or 10−7
m rhTSP-1 (E) before assaying for luciferase activity. Values are the mean ± S.E. from four experiments with duplicate determinations. C, activation of SMAD1 in C3H-B12 cells by ovine serum. C3H-B12 cells were exposed for 1 h to BMP-4 (10 ng/ml) or serum ½-diluted or serum ½-diluted supplemented with dorsomorphin (2.5 × 10−6
m). Total proteins were isolated from the cells, and Western blotting was performed with phospho-SMAD1 antibody. Equal loading of the proteins was confirmed by using GAPDH antibody. Bars with different letters indicate that group means are significantly different at p < 0.05. a.u., absorbance units.