Fig. 4

miR-155 promotes Phf19 expression by enhancing Akt signaling via downregulation of Ship1. a, b RNA-seq reads (a) and quantitative RT-PCR (b) of Phf19 mRNA in miR-155 and Ctrl-miR-overexpressing cells. Bars (mean ± s.e.m. of technical triplicates) are relative to Rpl13 mRNA. c Quantitative RT-PCR of Phf19 mRNA in in vitro activated KLRG1⁻ miR-155 sufficient and deficient CD8⁺ T cells. Bars (mean ± s.e.m. of technical triplicates) are relative to Rpl13 mRNA. d Ship1 and pAkt levels in miR-155-overexpressing cells assessed by Immunoblot. e pAkt levels of in vitro activated KLRG1⁻ miR-155 sufficient and deficient CD8⁺ T cells assessed by flow cytometry. f, g Quantitative RT-PCR of Phf19 mRNA in ex vivo sorted CD8⁺ T cells overexpressing miR-155 and Ctrl-miR after a 6 h incubation with or without AKT inhibitor VIII (Akti) (f) or transduction with constitutively active Akt (AktCA) or Thy1.1 control (g). Bars represent the mean ± s.e.m. of technical triplicates. h Ship1 and pAkt levels in Cas9⁺ CD8⁺ T cells transduced with Ship1-specific gRNA assessed by Immunoblot. i Quantitative RT-PCR of Phf19 mRNA in Cas9⁺ CD8⁺ T cells transduced with Ship1-specific gRNA or control. Bars represent the mean ± s.e.m. of technical triplicates. Data are representative of two independent experiments. *P < 0.05; **P < 0.01; ****P < 0.001 (unpaired two-tailed Student’s t-test)