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

FIGURE 4.

FIGURE 4.

Dominant-negative mutants of ALK1 and ALk2 inhibit BMP9 induced ALP activity in a dose-dependent manner. A and B, dnALK1 and dnALK2 inhibit BMP9 induced ALP activity in a dose-dependent fashion. Subconfluent C3H10T1/2 cells were infected with Ad-dnALK1, Ad-dnALK2 and/or Ad-RFP at three escalating titers, each of which had a 50% increase increment. At 24 h postinfection, cells were stimulated with BMP9-conditioned medium. ALP activity was measured at the indicated time points (A) and was stained histochemically (B) at day 7. Each assay condition was done in triplicate. C, dnALK1 inhibits BMP9 induced ALP activity in MEFs. Subconfluent MEFs were infected with Ad-dnALK1 and/or Ad-RFP at three escalating titers (50% increment). 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. D, dnALK2 inhibits BMP9 induced ALP activity in MEFs. Subconfluent MEFs were infected with Ad-dnALK2 and/or Ad-RFP at three escalating titers (50% increment). At 24 h post-infection, cells were stimulated with BMP9-conditioned medium. ALP activity was measured at the indicated time points. Each assay condition was done in triplicate.