Fig. 2. The anti pß-catenin antibody recognizes wild-type ß-catenin but not mutant ß-catenin or plakoglobin. (A) HA-tagged wild-type ß-catenin or mutant S33Y and S37A-ß-catenin were transfected into 293T cells and analyzed by western blotting with either anti-HA or pß-catenin antibodies. (B) 293T cells were transfected with HA-ß-catenin or with VSV-ß-catenin and HA-F-ß-TrCP. Immunopecipitation was performed with an anti-HA antibody and western blotting with pß-catenin or general ß-catenin antibodies. (C) 293T cells were treated with 30 mM LiCl overnight or with 25 µM MG132 for 4 hours and analyzed by western blotting for pß-catenin and total ß-catenin content. (D) VSV-tagged plakoglobin or ß-catenin were transfected into 293T cells, and their reactivity with pß-catenin antibody was determined when transfected alone or together with HA-tagged F-ß-TrCP. IP, immunoprecipitation; IB, immunoblot.