Fig. 3

HSP90AA1 is associated with FBXL6. a A partial list of proteins identified by mass spectrometry analysis. 293 T cells transfected with Flag-FBXL6 or Flag-Con plasmids were subjected to Flag M2 resin purification. Bound proteins were resolved by SDS-PAGE and stained with Coomassie blue staining, followed by mass spectrometry. b 293 T cells transfected with Flag-FBXL6 or Flag-Con plasmids were subjected to Flag M2 purification. Bound proteins were analyzed by immunoblot with indicated antibodies. c 293 T cells were infected with retroviruses encoding the indicated Flag-tagged F-box proteins (FBPs) and subjected to Flag M2 resin purification. The immunocomplexes were probed with HSP90AA1, SKP1 and Flag antibodies. Lane 1 shows a whole cell lysate from cells infected with an empty virus (EV). d The cell lysate of Hep3B cells were subjected to immunoprecipitation with anti-HSP90AA1 antibody or IgG. The bound proteins were detected by immunoblotting with antibodies against HSP90AA1 and FBXL6. e The cell lysate of SMMC-7721 cells were subjected to immunoprecipitation with anti-FBXL6 antibody or IgG. The bound proteins were detected by immunoblotting with antibody against HSP90AA1 and FBXL6. f 293 T cells were transfected with the indicated Flag-BRAT1 WT or deletion constructs for 36 h. Cells were harvested and lysed. The WCL was immunoprecipitated by anti-Flag M2 resin and immunoblotted with antibody against HSP90AA1 and Flag