Fig. 1.

OTUD1 regulates MCL1 protein stability. a Different cancer cell lines were treated with the proteasome inhibitor MG132 (10 µM) for 6 h. The cells were then harvested and subjected to SDS-PAGE and analysed by immunoblotting (IB) with the indicated antibodies. b Individual DUB expression plasmids were transfected into HeLa cells. Then, 36 h later, the cells were harvested and subjected to SDS-PAGE and analysed by immunoblotting with the indicated antibodies. c The empty vector or a plasmid expressing OTUD1 was transfected into Huh7 cells or MCF7 cells. Then, 36 h later, whole cellular extracts were subjected to Western blotting. d MCL1 RNA levels were determined in HeLa cells transfected with the empty vector or a plasmid expressing OTUD1. NS: no significant difference. e HeLa cells were infected with lentivirus expressing scramble shRNA or OTUD1-shRNA. Then, the cells were harvested and subjected to SDS-PAGE and analysed by immunoblotting with the indicated antibodies. f The MCL1-HA-expressing plasmid was co-transfected with the empty vector or a plasmid expressing OTUD1 into MCF7 cells. Then, 24 h later, the cells were treated with CHX for the indicated times. Next, the cells were 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