Fig. 5.

Both the TPR and U-box domains are required for MET degradation. (A) MET and GFP protein levels were determined by immunoblotting with GFP and MET antibodies, respectively. GFP was used as a control for efficient transfection and SDS-PAGE loading. (B) Cells were treated with MG132 and harvested. Lysates were analyzed by western blotting with antibodies for MET, GFP, and β-actin. (C) The cell lysates were immunoprecipitated using a polyclonal anti-MET antibody. The immunoprecipitation reactions were immunoblotted with a monoclonal anti-ubiquitin antibody (upper panel) and the corresponding cell lysates were probed using anti-MET. TPR, tetratricopeptide repeat; MET, mesenchymal-epithelial transition factor; GFP, green fluorescent protein; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; IP, immunoprecipitation analysis; IB, immunoblotting analysis; Ub, ubiquitination.