FIGURE 10.

Species differences in AR N/C interaction inhibition of AF2. A, CV1 cells were transfected with 3 μg of MMTV-Luc and 0.1 μg of human pCMV-hARΔ120–472 WT or L26A,F27A (LFAA), S94A (SA94), V33A (VA33), or K720A (KA720) mutant with 1.5 μg of pSG5 empty vector (−) and 2 μg of pSG5-TIF2 (T) with and without 1 μg of pSG5-MAGE (M). Cells were incubated in the absence and presence of 1 nm DHT. Inset, upper panel, pCMV5 empty vector (p5) (−) and pCMV-hARΔ120–472 WT and L26A,F27A, V33A, K720A, and S94A mutants (5 μg) were expressed in COS cells in the presence of 10 nm DHT. Lower panel, pCMV5 empty vector (−); pCMV-hARΔ120–472 WT, Q5, and S94A; and pCMV-mARΔ101–452 (5 μg) were expressed in COS cells. Cell extracts (60 μg of protein/lane) were analyzed by immunoblot using AR32 antibody. B, CV1 cells were transfected with 3 μg of MMTV-Luc and 0.1 μg of corresponding WT mouse pCMV-mARΔ101–452 or L26A,F27A, A33V, K700A, or A33V,K700A mutants with 1.5 μg of pSG5 empty vector (−) and 2 μg of pSG5-TIF2 (T) with or without 1 μg of pSG5-MAGE (M). Cells were incubated in the absence and presence of 1 nm DHT. Inset, pCMV5 empty vector (p5) (−) or pCMV-mARΔ101–452 WT, L26A,F27A, A33V (AV33), K700A (KA700), and A33V,K700A mutants (5 μg) were expressed in COS cells incubated with 10 nm DHT. Cell extracts (50 μg of protein/lane) were analyzed by immunoblot using AR32 antibody.