Figure 6.

TR4 repression of AR-mediated transcriptional activity. (A) Five hundred nanograms of MMTV-Luc was cotransfected with 40 ng of pCMV-AR (lanes 2 and 3) with increasing amounts of pCMX-TR4 (400, 800, and 1,200 ng). (B) Five hundred nanograms of PSA-Luc was cotransfected with 40 ng pCMV-AR (lanes 2 and 3) with increasing amounts of pCMX-TR4 (400 and 1,200 ng). After 24 hr transfection, cells were treated with 10 nM of DHT. After 16–18 hr incubation, cells were harvested for dual-luciferase reporter assay. (C) Northern blotting analysis of PSA transcripts in LNCaP cells. Total RNA (25 μg) from LNCaP cells, which were transfected with either pCMX-TR4 or pCMX vector by using SuperFect (Qiagen), was applied into a formamide RNA gel, transferred onto a nylon membrane, and hybridized with a ³²P-PSA gene fragment from the exon 3. β-actin was used as an internal control.