Effects of BFA on cell cycle. (A) LNCaP cells were treated with DHT (1 nM), BFA (30 ng/ml), or BFA/DHT combination for 72 h and cell cycle analysis was performed as described in Methods. Cell cycle phase distributions or the number (%) of cells present at the G1, S, or G2/M phases in each experimental condition was determined and plotted. The data were mean ± SD from three separate experiments and subjected to statistical analysis; however, only those mean values (without SD) were used for plotting the graph for a clear illustration. (B) After cells were treated with or without BFA (30 ng/ml) for 72 h, the expressions of specific cell cycle regulators such as CDK2, CDK4, cyclin D1, and p21WAF1 were analyzed on Western blots and autoradiographs of those regulators (on the X-ray film) are shown.