Fig. 2

OTUD1 regulates MCL1 protein stability in an enzymatic activity-dependent manner. a The indicated cells were infected with lentivirus expressing empty vector, OTUD1 or its enzymatic activity mutant C320S. Then, 36 h later, whole cellular extracts were subjected to Western blotting. b HeLa cells were infected with lentivirus expressing empty vector, OTUD1 or its functional domains. Then, 36 h later, whole cellular extracts were subjected to Western blotting. c The MCL1-HA-expressing plasmid was co-transfected with a plasmid expressing OTUD1 or its enzymatic activity mutant C320S into HeLa cells. Then, 24 h later, the cells were treated with CHX for the indicated times. The cells were then harvested and subjected to SDS-PAGE and analysed by immunoblotting with the indicated antibodies. The intensities of the MCL1-HA expression levels were quantified by densitometry and plotted. The experiment was repeated three times, and a representative experiment is presented. **p < 0.01