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. 2022 Mar 16;29(9):1864–1873. doi: 10.1038/s41418-022-00971-8

Fig. 2. OTUB1 enhances MYC protein stability.

Fig. 2

Immunoblotting analyses were performed with the indicated antibodies. A 293 T cells were overexpressed HA-tagged MYC and Flag-tagged OTUB1 (WT or mutants) proteins. Immunoblotting analyse was performed. B 293 T cells were overexpressed HA-tagged MYC and Flag-tagged OTUB1 (0,0.5 or 1 μg) proteins. Immunoblotting analyse was performed. C, E MCF-7 or MB-231 cells were knocked down OTUB1 with shRNA. Immunoblotting analyse was performed. D, F MCF-7 or MB-231 cells were overexpressed Flag-tagged OTUB1 proteins. Immunoblotting analyse was performed. G MCF-7 or MB-231 cells were knocked down OTUB1 with shRNA. Immunofluorescence was performed. H MCF-7 cells with overexpression Flag-tagged OTUB1 protein were treated with MG132 for 8 hr. Immunoblotting analyse was performed. I 293 T cells were overexpressed the indicated HA-tagged MYC and Flag-tagged OTUB1 or OTUB2 proteins. Immunoblotting analyse was performed. J MCF-7 cells with overexpression of Flag-tagged OTUB1 were treated with CHX for indicated time. Immunoblotting analyse was performed. K MCF-7 cells with knockdown of OTUB1 were treated with CHX for indicated time. Immunoblotting analyse was performed. L MEFs cells with knockout of OTUB1 were treated with CHX for indicated time. Immunoblotting analyse was performed.