Figure 10

LMP1 regulated the ubiquitination of SENP2. (a) EBV-transformed LCLs were grown and denaturing immunoprecipitations (to pull-down all proteins covalently modified by endogenous ubiquitin) were performed. (b) HEK 293 cells were transfected and denaturing immunoprecipitations were performed to pull-down all proteins covalently modified by exogenous HA-ubiquitin (HA-Ub). Western blot analyses were done to detect SENP2 (endogenous or GFP), LMP1, and/or myc-SUMO1 levels. (c,d) HEK 293 cells were transfected as indicated and treated with PYR-41 or the vehicle control (DMSO) for 18 hours prior to harvesting. (c) WCL were collected and Western blot analyses performed to detect endogenous ubiquitin levels. GAPDH was used as the loading control. (d) Denaturing immunoprecipitations were performed to pull-down all proteins covalently modified by exogenous HA-ubiquitin (HA-Ub). Western blot analyses were performed to detect SENP2 or HA-ubiquitin.