Fig. 1. Evaluation of rhBMP-2 efficacy. C2C12 cells were cultured and treated with rhBMP-2 obtained from two different sources (R & D Systems, R&&D, and joint Protein Central, jPC). (A) Osteogenic efficacies were similar, as determined by ALP staining. The intensity of each well was quantified by an image analysis program. Bars indicate the relative intensity ratio of experimental groups after normalization with the control group (right panel, n = 3). (B) rhBMP-2 increased Smad1,5,8 phosphorylation and Runx2 expression. Cells were cultured and treated with various concentrations of rhBMP-2 described in the Materials and Methods section. Total proteins were extracted and determined by phosphorylation of Smad1/5/8 (P-smad 1/5/8, left panel) and Runx2 expression (right panel) by western blot analyses. Smad1 and β-actin were loading controls.
