Figure 2

TGFβ promotes cyclin D1 nuclear accumulation and co-localization with p21. (A) MDA and SCP2 cells were treated with or without 5 ng/ml transforming growth factor beta (TGFβ) for 24 hours. Immunocytochemistry was performed using cyclin D1 (red) antibody and DAPI (blue). The scale bar is 10 µm. (B) MDA cells were immunostained using p21 (green) and cyclin D1 (red) antibodies. Co-localizaton (yellow) between p21 and cyclin D1 is represented by overlay. (C) MDA and SCP2 cells were treated with TGFβ for the indicated times. Cell lysates were analyzed by co-immunoprecipitation using an anti-cyclin D1 antibody. Immunoprecipitated cyclin D1 was subjected to Western blotting. (D) MDA, SUM149 and SUM159 cells were treated with TGFβ. Cell lysates were immunoprecipitated using an anti-p21 antibody and analyzed by immunoblotting using anti-cyclin D1 and anti-p21 antibodies.