Fig. 7.

The ability of a DNA-binding mutAR to support proliferation in LP50 cells. A, LP50 cells were nucleofected with AR shRNA and cotransfected with either mutAR or the vector control (Ctrl). Cells were also cotransfected with the control shRNA plasmid and the vector control. Cells were harvested 72 h after transfection for Western blot analysis or to extract total RNA. The Western blotting was probed for AR and for the GAPDH loading control (A, inset). The RNA was reverse transcribed, and the mRNAs for endogenous AR and GAPDH were measured by real-time PCR. A, The endogenous AR was distinguished from that of mutAR by targeting the TaqMan probe to its 5′UTR. B, In the same experiment as in A, 72 h after transfection, [³H]thymidine incorporation was measured in the cells in separate sets of triplicate wells.