Fig. 6

Phf19 is a critical downstream factor of miR-155 in CD8⁺ T cells. a Venn diagrams and b volcano plot depicting the number of overlapping differentially expressed genes between differentially expressed genes in miR-155-overexpressing and Phf19^−/− KLRG1⁻CD62L⁻ CD8⁺ T cells. Gene expression was evaluated by RNA-seq of pmel-1 KLRG1⁻CD62L⁻ CD8⁺ T cells 5 days after adoptive transfer of 3 × 10⁵ pmel-1 Phf19^+/+/Phf19^−/− cells or pmel-1 cells overexpressing Ctrl-miR/miR-155 into wild-type mice in conjunction with gp100-VV. RNA-seq data were obtained from triplicated groups of three individual mice. c Gene sets significantly enriched (FDR < 0.25) in Phf19^−/− CD8⁺ T cells. Gene sets also enriched in miR-155 overexpressing cells are highlighted in black. d Enrichment of genes upregulated in CD8⁺ T cells responding to primary vs secondary LCMV infections⁴³ (left panels) and enrichment of genes upregulated in CD8⁺ T cells at d6 vs d10 post LmOVA infections⁴⁵ (right panels) in miR-155-overexpressing and Phf19^−/− CD8⁺ T cells. e Flow cytometry analysis of congenically-distinguishable live pmel-1 Phf19^+/+ Ly5.2^+/+ and pmel-1 Phf19^−/− Ly5.1^+/− cells transduced with either miR-155 or Ctrl-miR assessed pre-transfer and 5 days after co-transfer of 3 × 10⁵ cells into wild-type mice in conjunction with gp100-VV administration. Numbers adjacent to outlined areas indicate percentage after gating on live CD8⁺GFP⁺ T cells. f, g Percentages of live pmel-1 CD8⁺GFP⁺ T cells transduced with Ctrl-miR (left panel) or miR-155 (right panel) (f) and CD8⁺ GFP⁺ T_E cells (g) at indicated time points after transfer as described in e. Symbols represent the mean ± s.e.m. of three individual mice; small horizontal lines (right panel) indicate the mean ± s.e.m. Data are representative of two independent experiments. ***P < 0.005; ****P < 0.001 (unpaired two-tailed Student’s t-test)