MDA-MB-231 cells were transfected with scrambled, CCND1(51), and/or CCND1(52) siRNA. The amount of cyclin D1 protein was lower after transfection with the cyclin D1-specific siRNA compared to scrambled siRNA. The amount of CDK4 and FLNa was not affected while the amount of pFLNa (ser-2152) (A) and pFLNa(ser-1459) (B) decreased in cells transfected with the cyclin D1-specific siRNA with greater effect with CCND1(51).
Filamin A (FLNa) is a binding partner of cyclin D1. (C) Anti-cyclin D1 immunoprecipitations were performed using MDA-MB-231 protein lysate. Phosphorylated FLNa was detected by Western blot of the immunoprecipitated fraction. (D) Anti-FLNa immunoprecipitations were performed using MDA-MB-231 protein lysate. Cyclin D1 was detected by Western blot of the immunoprecipitated fraction.