Fig. 7. BMP2 stimulates LIMK1 activity. (A) Serum-starved C2C12 cells were stimulated with 3 nM BMP2 and cell lysates analyzed by immunoblotting with anti-LIMK1-P and anti-LIMK1 antibody. The bottom panel indicates the relative LIMK1-P levels. Mean ± s.e.m. from three independent experiments (*P<0.05, paired t-test). (B) C2C12 cells were treated as above for 40 minutes but in the absence or presence of LY294002. Cell lysates were analyzed by immunoblotting with the indicated antibodies. The bottom panel indicates the relative LIMK1-P levels. Mean ± s.e.m. from three independent experiments (*P<0.05, paired t-test). (C) C2C12 cells were treated with BMP2 and LY294002 as indicated. Endogenous LIMK1 was subjected to an in vitro kinase assay with GST-cofilin as a substrate. Values of cofilin phosphorylation with respect to the LIMK1 present in cell extracts are shown.