Figure 2.
RB depletion confers no proliferative advantage in the presence of androgen. A, LNCaP cells transiently transfected with pMSCV-puroRb3C or pMSCV-puro (hashed columns) or stable clones described in Fig. 1B, shRB1 and shCon1 (solid columns) were cultured in ΔFBS and monitored for BrdUrd incorporation as described in Materials and Methods. For comparison, BrdUrd incorporation in each RB control was set to 1. B, shRB1 and shCon1 cells were seeded at equal density in the presence of androgen (ΔFBS), and after 24 h of culture (time, 0 h), cell number was monitored at the times indicated. Data represent at least three independent experiments wherein data for each time point were collected with triplicate biological replicates (P > 0.05, ANOVA). C, asynchronous populations of shRB1 and shCon1 cells were cultured for 48 h, pulsed with BrdUrd for 1 h, and then analyzed for cell cycle and BrdUrd incorporation. Data represent four independent samples (P > 0.05, ANOVA).