Figure 2.

GTIN modulated cell-cycle regulatory factors in VSMCs. A7r5 cells were incubated with GTIN at the indicated doses (0, 1, 5 and 10 μM) for 48 h. The protein levels of p-p53, p53 and CDKIs, including p27, p21 and p16 (a), E2F1, Rb, p-Rb (b) and β-actin, as an internal control, were assayed by WB. (c) The expressions of Rb/E2F1, cyclin D1/cdk-4 and cyclin E/cdk-2 complexes were determined. The protein lysate was immunoprecipitated (IP) with anti-E2F1, cdk-4 or cdk-2 antibodies. The precipitated complexes were then analyzed for immunoblotting (IB) applying antibodies against Rb, cyclin D1 or cyclin E. All quantitative results were showed as mean ± SD of three repeats from at least three independent experiments. * p < 0.05, ** p < 0.01 compared with the untreated control via one-way ANOVA with Dunnett’s post-hoc analysis.