Fig. 2

Ezh2 is required for the enhanced CD8⁺ T cell antitumor immunity conferred by miR-155. a Number of live CD8⁺GFP⁺ T cells in the spleen assessed at day 5 and 10 following adoptive transfer of 3 × 10⁵ pmel-1 Ezh2^+/+ or Ezh2^−/− CD8⁺ T cells transduced with miR-155 or Ctrl-miR into B16 tumor-bearing mice in conjunction with gp100-VV. Bars represent the mean ± s.e.m. of three or two individual mice. b Flow cytometry of splenic CD8⁺GFP⁺ T cells 5 and 10 days after transfer as in a. Numbers indicate the percentage of cells after gating on live CD8⁺GFP⁺ T cells. c Percentage of splenic pmel-1 CD8⁺GFP⁺ T_E cells at indicated time points after transfer. Data are presented as box plots extending to minimum and maximum values. Bands inside the boxes represent median values of three individual mice. d Combinatorial cytokine response of splenic CD8⁺GFP⁺ T cells 5 and 10 days after transfer as in a, as determined by the Boolean combination of gates identifying IFN-γ⁺, IL-2⁺, and TNF⁺ cells. Data are presented as the mean of three individual mice. e−f Tumor size (mean ± s.e.m.) (e) and survival curve (f) of B16 tumor-bearing mice after adoptive transfer of 4 × 10⁶ cells generated as in a in conjunction with gp100-VV (n = 5 mice/group). NT no treatment. Data are representative of two independent experiments. *P < 0.05; **P < 0.01 (unpaired two-tailed Student’s t-test) (a, d), **P < 0.01 [a Log-rank (Mantel-Cox) Test] (f)