Figure 3.

Effects of isorhamnetin on the levels of cell cycle regulatory proteins in bladder cancer cells. (A) T24 and 5637 cells were treated with the indicated concentrations of isorhamnetin for 48 h, and then total cell lysates were prepared. Western blotting was then performed using the indicated antibodies and an enhanced chemiluminescence (ECL) detection system. Actin was used as an internal control. (B) The expression of each protein was indicated as a fold change relative to the control. Quantitative analysis of mean pixel density was performed using the ImageJ^® software. (C) Cells were incubated without or with 100 μM isorhamnetin for 48 h, and then equal amounts of proteins were immunoprecipitated with the anti-cyclin-dependent kinase (Cdk) 1 antibody. Western blotting using immunocomplexes was performed using anti-p21 or anti-Cdk1 antibodies and an ECL detection system (IP, immunoprecipitation).