Fig. 1.

Role of androgen and AR in the proliferation of EP-LNCaP and LP50 cells. A, Hormone-depleted LP50 cells were treated with R1881 (1 nm) or vehicle (veh). The cells were counted in a coulter particle counter. B, EP-LNCaP and LP50 cells were hormone depleted. The endogenous AR protein levels or GAPDH [loading control (ctrl)] were measured by Western blotting (inset). The mRNA for AR and PSA were measured by real-time RT-PCR. C, Hormone-depleted EP-LNCaP cells were treated with R1881 (1 nm) or vehicle. At 18 h of the treatment, [³H]thymidine was added to the media, and 6 h later, the incorporation of the radiolabel into DNA was measured. D, LP50 cells were infected with AR shRNA lentivirus or nontarget control lentivirus; 12 h after infection, the cells were treated to deplete them of hormone and to measure [³H]thymidine incorporation in response to treatment with either vehicle or R1881, all exactly as described for C. D, inset, Western blotting showing the expression of AR or GAPDH (loading control) in LP50 cells infected with AR shRNA lentivirus or nontarget control shRNA lentivirus and treated with vehicle or R1881. CPM, Count per minute; *, P < 0.001.