FIG. 5.

RNF31 stabilizes endogenous DAX-1. α, anti. (A) H295R cells were treated with 10 μM proteasome inhibitor MG132 for 6 h and analyzed by Western blotting with indicated antibodies. (B) H295R cells transfected with either Flag or Flag-RNF31 were treated with cycloheximide (CHX) 24 h posttransfection. Cells were lysed 0, 1, 3, 6, 9, and 24 h after CHX addition, and extracts were analyzed by Western blotting using indicated antibodies. (C) H295R cells were treated with either control (−) or RNF31-specific (+) siRNA. Endogenous proteins from whole-cell extracts were analyzed by Western blotting using indicated the antibodies. Results of independent transfections of each oligonucleotide are shown.