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. 2024 Aug 14;300(9):107672. doi: 10.1016/j.jbc.2024.107672

Figure 4.

Figure 4

USP7 deubiquitinates ZMYND8.A, immunoblots of WCL and co-IP from HEK293T cells transfected with Flag-ZMYND8 vector, WT or single-lysine only HA-ubiquitin (Ubi) vector, or Myc-USP7 vector and treated with 10 μM MG132 for 6 h before harvest. B, ubiquitination of endogenous ZMYND8 in HEK293T ectopically expressing WT or C223S Myc-USP7. C and D, ubiquitination of endogenous ZMYND8 in MDA-MB-231 cells (C) and MDA-MB-231 cells (D) stably expressing Myc-USP7. E and F, ubiquitination of endogenous ZMYND8 in SC and USP7-KD MDA-MB-231 cells (E) and MDA-MB-231 cells (F). G, ubiquitinated Flag-ZMYND8 was precipitated from HEK293T cells transfected with constructs encoding Flag-ZMYND8 and HA-Ubi-K48 and incubated with purified WT or C223S Myc-USP7 in deubiquitylation buffer for examination of ZMYND8 ubiquitination. H, ubiquitination of endogenous ZMYND8 in MDA-MB-231 cells stably expressing Myc-FBXW7 with or without Myc-USP7.