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. 2022 Mar 6;18(5):2186–2201. doi: 10.7150/ijbs.60594

Figure 6.

Figure 6

CSN5 implicate ZEB1 ubiquitination. The MDA-MB-231 cells were transfected with oe-Vector plasmid or oe-CSN5 plasmid by lentiviral vector. A The efficiency of CSN5 overexpression were measured by qPCR. ShRNA-NC-MDA-MB-231 cells, shRNA-SENP1-MDA-MB-231 cells were transfected with oe-Vector plasmid or oe-CSN5 plasmid by lentiviral vector. B And the cell extracts from the indicated groups were subjected to immunoprecipitation with anti-ZEB1 antibody or anti-IgG antibody, followed by immunoblot with anti-ubiquitin antibody. C, D, E, F, G The expression of E-cadherin, Krt12, Vimentin, Survivin in different group were detected by western blot assay. MDA-MB-231 cells were transfected with ShRNA-NC+Flag-ZEB1-WT, ShRNA-CSN5+Flag-ZEB1-WT, ShRNA-NC+Flag-ZEB1-K1108A, ShRNA-CSN5+Flag-ZEB1-K1108A, ShRNA-NC+Flag-ZEB1-K186A, ShRNA-CSN5+Flag-ZEB1-K186A, ShRNA-NC+Flag-ZEB1-K439A or ShRNA-CSN5+ Flag-ZEB1- K439A by lentiviral vector. H The cell extracts from the indicated groups were subjected to immunoprecipitation with anti-ZEB1 antibody or anti-IgG antibody, followed by immunoblot with anti-ubiquitin antibody. Results were mean ± SD for three individual experiments which, for each condition, were performed in triplicate. * P <0.05, ** P <0.01.