Figure 5: Exogenous expression of EZH2^Y641F in lymphocytes improves tumor killing and adoptive T cell therapy.

(A) Western blot analysis of in vitro activated CD8⁺ wt and Lck-EZH2^Y641F T cells. (B) Oxygen consumption rate of pre-activated CD8⁺ wt and Lck-EZH2^Y641F T cells. P value was determined by unpaired t test. (C) In vitro killing assay was used to determine cytotoxic function. Target cells (MC38 ^SIINFEKL) were cultured with CellTrace-Violet labeled, pre-activated OT-1 or OT-1- EZH2^Y641F CD8+ T cells at a target/effector ratio of 1:0, 1:1, or 1:16. Tumor killing is presented as the percentage of viable (Annexin V⁻, PI⁻) tumor cells remaining after 10 hours of co-culture. Data are representative of 2 independent experiments. (D) C57BL/6 mice were inoculated with 5×10⁶ MC8^SIINFEKL. After 9 days 4×10⁶ WT, OT-I, or OT-I. EZH2^Y641F pre-activated CD8⁺ T cells were transferred I.V. into recipients and tumor growth was assessed. Tumor growth curves depict an average tumor volume in each group, n=8–10 and error bars indicate the SEM. Asterisk denotes statistical significance P<0.01.