Fig. 3

Involvement of p53 in compound C1–induced G2/M arrest. a A549 cells were treated with C1 for 12 h and harvested for western blotting of p53 and p21 and cell cycle analyses. As a positive control, A549 cells were irradiated with 10 Gy X-ray and harvested 12 h after irradiation for Western blotting analyses of p53 and p21; actin was used as a loading control. Representative blots of two different experiments are shown [upper panel]. Representative histograms of two different experiments are shown [lower panel]. b A549 cells preincubated with the caffeine were cultured in the presence of 10 μM C1 for 24 h [left panel] or cultured for 12 h after 10 Gy-irradiation [right panel]. The cells were harvested, and cell cycle analysis was performed. Representative histograms of two different experiments are shown. c–d A549 cells treated with p53 siRNA were cultured in the presence of C1 at 10 μM for 12 h, and then western blotting of p53 (c) and cell cycle profiles (d) were analyzed. Representative results of two different experiments are shown. e A549 cells were treated with C1 for 12–24 h and harvested for western blotting of phospho-cdc2 (p-cdc2), cyclin B1, and phospho-histone H3 (p-H3). Representative results of two different experiments are shown