Figure 5.

BMP9 induces osteogenic differentiation in iMADs in vitro. (A) BMP9 effectively induces osteogenic marker alkaline phosphatase (ALP) activity in iMADs. Subconfluent iMADs were infected with Ad-BMP9 or Ad-GFP. At 5 days after infection, cells were fixed for histochemical staining for ALP activity (a). Quantitative measurement of relative ALP activity was also determined at 3, 5, and 7 days after infection (b). Assays were done in triplicate and representative images are shown. “**”, p<0.001. (B) BMP9 induces robust matrix mineralization. Ad-BMP9 or Ad-GFP infected iMADs were cultured in mineralization medium for 10 days and stained with Alizarin Red S. Assays were done in triplicate and representative images are shown. (C) BMP9 has limited effect on adipogenic differentiation, Ad-GFP (a) or Ad-BMP9 (b) infected iMADs were cultured in complete DMEM. At 10 days after infection, the cells were fixed and stained with Oil Red O solution. Assays were done in triplicate and representative images are shown.