Fig. 6: OTUD7B deubiquitylates ERα.
From: OTUD7B stabilizes estrogen receptor α and promotes breast cancer cell proliferation
A MCF-7 cells transfected with the indicated siRNA were treated with MG132 for 6 h before collection. ERα was immunoprecipitated with anti-ERα and immunoblotted with anti-HA. B Immunoblotting to detect the ubiquitination of ERα in HEK293 cells cotransfected with Flag-ERα, HA-Ubiquitin, and Myc-OTUD7B (wild type or C194S). C Immunoblotting to detect the ubiquitination of ERα in HEK293 cells cotransfected with Flag-ERα, HA-Ubiquitin, and Myc-OTUD7B (wild type or C194S). D OTUD7B removed the ubiquitin chain of ERα in a dose-dependent manner. E OTUD7B removed the ubiquitin chain of ERα in a time- and dose-dependent manner. F ERα ubiquitylation was analyzed in cells transfected with E3 TRIM8 together with OTUD7B or not. G HA-WT, K6, K11, K27, K29, K33, and K48 or K63 Ub was cotransfected with Flag-ERα and Myc-OTUD7B into HEK293 cells. After treatment with 10 μM MG132 for 6 h, cell lysates were subjected to ubiquitination assay and the ubiquitination level of ERα was detected by HA antibody. *P value < 0.05, **P value < 0.01, ***P value < 0.001.
