Figure 3.

FBW7-mediated degradation of EZH2 is Thr²⁶¹-dependent. A, Western blotting analysis of co-immunoprecipitation of ectopically expressed FLAG-EZH2 WT and FLAG-EZH2 T261A and Myc-FBW7α in 293T cells. Data are representative of multiple experiments (n = 3). IB, immunoblot. B, PANC-1 cells were transfected with the indicated plasmids for 24 h, followed by Western blotting analysis. Data are representative of multiple experiments (n = 3). C and D, PANC-1 cells were transfected with the indicated plasmids. After 24 h, cells were treated with 50 μg/μl CHX. At different time points, cells were harvested for Western blotting analysis. E, PANC-1 cells were transfected with the indicated plasmids for 16 h, followed by treatment with 20 μm MG132 for 8 h. Immunoprecipitated FLAG-EZH2 proteins were analyzed by WB for ubiquitination.