Fig. 6

FAM83H directly bind to β-catenin and regulates the stabilization of β-catenin protein by the proteasome-mediated ubiquitination of β-catenin. a Immunoprecipitation for FAM83H and β-catenin. β-catenin was detected in the protein taken with immunoprecipitation for FAM83H and vice versa. b Western blot for FAM83H, β-catenin, actin, and lamin B1 with the protein lysates fractionated according to subcellular localization after transfection of shRNA for FAM83H in MG63 cells. c Confocal microscopic analysis with immunofluorescence staining for FAM83H (red) and β-catenin (green) after knock-down of FAM83H with shRNA for FAM83H in MG63 cells. The co-localization of FAM83H and β-catenin is colored yellow. d The MG63 cells were transfected with control vector or shRNA for FAM83H. 48 h after transfection, the cells were treated with 20 μg/ml cycloheximide or 20 μM MG132 for 0.5 to 2 h. The cell lysates were blotted with anti-β-catenin antibody and anti-β-actin antibodies. e The MG63 cells were transfected with control vector or shRNA for FAM83H for 48 h. Subsequently, the transfected cells were treated with MG132 (20 μM) for 4 h and total lysates of cells were immunoprecipitated with anti-β-catenin antibodies and blotted with anti-ubiquitin and anti-β-catenin antibodies