FIG. 11.

The N-CoR interacts with the N-terminal domain of the AR and inhibits the AR-AF-1-mediated transcriptional activation. (A) The N-CoR interacted with the N-terminal domain of the AR. COS-7 cells were cotransfected with expression plasmids for AR-AF-1 or AR-AF-2 and VP16 or VP16-fused N-CoR mutants with pGL3-MMTV and pRL-CMV for the mammalian one-hybrid assay. The molar ratio of the transfected amount of expression plasmids for AR-AF-1 or AR-AF-2 and N-CoR was 1:5. The cells were incubated for 24 h in the absence (for AR-AF-1) or presence (for AR-AF-2) of 10 nM DHT. The luciferase activities are represented as values relative to the activity induced by AR-AF-1 alone in the absence of ligand. (B) Inhibition of AR-AF-1-mediated transactivation by the N-CoR. COS-7 cells were cotransfected with expression plasmids for AR-AF-1 and the full-length N-CoR or its mutants with pGL3-MMTV and pRL-CMV. The cells were incubated for 24 h in the absence of DHT. The luciferase activities are represented as values relative to the activity induced by AR-AF-1 alone in the absence of ligand. Each bar in panels A and B represents the mean ± standard deviation of results from three independent experiments. *, P < 0.05. (C) Subcellular distribution of AR-AF-1-YFP in COS-7 cells. (D) Redistribution of GFP-N-CoR by nonfluorescent AR-AF-1 in the absence of DHT. An expression plasmid for GFP-N-CoR (i), GFP-N-CoR(1-1798) (ii), GFP-N-CoR(1-1133) (iii), GFP-N-CoR(1134-1798) (iv), or GFP-N-CoR(1803-2439) (v) was cotransfected into COS-7 cells with the plasmid for nonfluorescent AR-AF-1. Bars, 5 μm.