Fig. 7

Phf19 epigenetically reprograms T cell antitumor immunity. a Amino acid sequence alignment of the partial aromatic cage of Phf19. Tryptophan (W) and tyrosine (Y) residues were mutated to cysteine (C) or alanine (A), respectively. b Immunoblot of total, soluble, and chromatin-bound proteins from CD8⁺ T cells transduced with the Phf19Thy1.1, Phf19mutThy1.1, and Thy1.1 control. c Flow cytometry of splenic T cells at indicated time points following adoptive transfer of 3 × 10⁵ pmel-1 CD8⁺ T cells transduced with Phf19Thy1.1, Phf19mutThy1.1, or Thy1.1 into wild-type mice in conjunction with gp100-VV. Numbers indicate percentage after gating on live CD8⁺ Thy1.1⁺ T cells. d Number of splenic pmel-1 CD8⁺Thy1.1⁺ T cells 5 days after transfer as in c. Bars represent the mean ± s.e.m. of three individual mice. e Flow cytometry of splenic CD8⁺Thy1.1⁺ T cells at indicated time points after transfer as in c. Numbers indicate percentage after gating on live CD8⁺Thy1.1⁺ T cells. f Percentage of splenic pmel-1 CD8⁺Thy1.1⁺ T_E cells at indicated time points as in c. Data are presented as box plots extending to minimum and maximum values. Bands inside the boxes represent median values of three individual mice. g Intracellular cytokine staining of splenic CD8⁺Thy1.1⁺ T cells 5 days after transfer as in c. Numbers indicate percentage after gating on live CD8⁺Thy1.1⁺ T cells. h Combinatorial cytokine response of splenic CD8⁺Thy1.1⁺ T cells as in c, 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. i Tumor size (left, mean ± s.e.m.) and survival curve (right) of B16 tumor-bearing mice receiving 2 × 10⁶ cells generated as in c in conjunction with gp100-VV and IL-2 (n = 7 mice/group). NT no treatment. Data are representative of two independent experiments. *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001 (unpaired two-tailed Student’s t-test) (d, h, and i left); *P < 0.05 (a Log-rank (Mantel-Cox) Test) (i right)