EZH2 Y244A increases the interactions of EZH2 with PRC2 subunits and reduces the interaction with Pol II. (A) Coimmunoprecipitation assay shows the interaction between endogenous EZH2 and Pol II in NKTL cell lines. (B) Coimmunoprecipitation assay shows the interaction between endogenous Pol II and JAK3 in NKYS cells. (C) Coimmunoprecipitation assay shows the changes in the association of EZH2 WT /Y244A with Pol II, SUZ12, and EED. 293T cells were cotransfected with JAK3 A572V and EZH2 WT/Y244A expression plasmids or control vector for 48 hours. (D) ChIP-qPCR assay in 293T cells for binding of ectopically expressed EZH2WT/EZH2 Y244A to EZH2 target genes. ChIP assays were performed by using 293T cells cotransfected by JAK3 A572V and EZH2 WT/Y244A expression plasmids. Real-time PCR was performed with immunoprecipitated chromatin fragments obtained by using an anti-HA antibody. Results are means ± SD. n = 3 independent experiments. *P < .05; **P < .01 (Student t test). (E) qRT-PCR assay in 293T cells showing that EZH2-Y244A mutation affects EZH2 target gene expression. Results are means ± SD. n = 5 independent experiments. *P < .05; **P < .01 (Student t test).