FIG. 4.

The AR acetylation site regulates coactivator-mediated induction of ligand-treated AR. The MMTV-LUC reporter was cotransfected with expression vectors for either the wild-type or mutant AR and the candidate AR coactivators TIP60 (A), SRC1 (B), ARA55 (C), ARA70 (D), Ubc9 (E), and p300 (F) or equal amounts of the empty expression vector cassettes (pCMV). The cells were treated with either DHT (10⁻⁷ M) or vehicle for 24 h, and luciferase activity was assessed. The data are shown as the means ± standard deviations for at least six separate transfections. DHT treatment (10⁻⁷ M) was for 24 h. (G) The ARwt and AR acetylation site mutant were transfected into HEK293 cells, and Western blotting was performed for AR, p300, and the loading control guanine nucleotide dissociation inhibitor (GDI). In the right panel, equal amounts of the cellular extracts were immunoprecipitated with the AR antibody, and Western blotting was performed with either the AR or the p300 antibody. +, present; −, absent.