Additional file 3: Figure S3. of E2-EPF UCP regulates stability and functions of missense mutant pVHL via ubiquitin mediated proteolysis

Ubiquitination is decreased in lysine zero mutant pVHL in cells significantly. (C) Single-lysine (K159, K171, and K196) and lysine-zero mutant pVHLs were analyzed in vitro ubiquitination assay. GST-tagged VHL was incubated with E1, Flag-ubiquitin and His-UCP at 37 °C for 1 h. Each GST-VHL was pulled-down with glutathione sepharose beads and analyzed by anti-GST or anti-HA antibody. (TIFF 115 kb)