Skip to main content
. 2010 Jul 13;285(38):29588–29598. doi: 10.1074/jbc.M110.130518

FIGURE 3.

FIGURE 3.

Dominant-negative mutants of ALK1 and ALk2 inhibit BMP9 induced ALP activity in pre-osteoblast progenitor cells. A, dnALK1 and dnALK2 inhibit BMP R-Smad reporter activity induced by BMP9. Subconfluent C3H10T1/2 cells were transfected with 12xSBE-Luc reporter and infected with Ad-dnALKs or Ad-RFP. At 24 h post-transfection/infection, cells were stimulated with BMP9-conditioned medium. Luciferase activity was measured at the indicated time points. Each assay condition was done in triplicate. B and C, inhibition of BMP9 induced ALP activity by dnALK1 and dnALK2. Subconfluent C3H10T1/2 cells were infected with Ad-dnALKs or Ad-RFP. At 24-h postinfection, cells were stimulated with BMP9-conditioned medium. ALP activity was measured at the indicated time points (B) and was stained histochemically (C) at day 7. Each assay condition was done in triplicate. D, dnALK1 and dnALK2-mediated inhibition of BMP9 induced ALP activity in pre-osteoblast progenitor cells. Subconfluent C2C12 cells, MEFs and BMSCs were infected with Ad-dnALKs or Ad-RFP. At 24 h postinfection, cells were stimulated with BMP9-conditioned medium. ALP activity was measured at the indicated time points. Each assay condition was done in triplicate.