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. 2020 Sep 16;16(15):2951–2963. doi: 10.7150/ijbs.47999

Figure 8.

Figure 8

USP14 interacts with MDM2 and upregulates MDM2 protein level. (A) Immunofluorescent staining of GFP was executed after co-transfected with cherry-MDM2 and flag-USP14 plasmids for 24 h in HeLa cells. The co-localization between USP14 and MDM2 was revealed by immunofluorescent staining. (B) Co-immunoprecipitation (Co-IP) assay for the interaction between USP14 and MDM2 in HeLa cells. Total proteins were extracted from HeLa cells, immunoprecipitated with USP14 antibody beads and immunoblotted for MDM2. (C-F) After transfected with pLVX-puro vector or pLVX-USP14-HA for 48 h, the levels of HA (USP14) and MDM2 protein were detected by western blot in HeLa cells. (G-J) The HeLa or 293T cells were transfected with pLKO.1-shUSP14 or pLKO.1-EGFP-Puro vector (scramble shRNA control) for 48 h, and then the levels of USP14 and MDM2 protein were measured by western blot. *p < 0.05; **p< 0.01.