Panel A shows a schematic diagram of retroviral transduction and adoptive transfer experiment. Panel B shows line graph comparing tumor growth among experimental groups. Panel C shows individual replicate tumor growth curves for each group. Panel D shows tumor weight and infiltrating O T I cells. Panel E shows flow cytometry plots and quantification of G F P positive O T I cells. Panel F shows flow cytometry plots and quantification of T c f 1 positive T P E X and T E X cells. Panel G shows flow cytometry plots and quantification of C X 3 C R 1 positive intermediate T E X cells. Panel H shows flow cytometry plots and quantification of G z m B, I F N gamma, and Perforin expression.
Reconstitution of Mettl8 expression in Mettl8 −/− OT-I cells restored their phenotype to that of WT OT-I cells. (A) Schematic diagram of the rescue experiment: CD45.2 mice were subcutaneously injected with 2 × 105 EG7-OVA cells. Mettl8 overexpression (OE) or empty vector (EV) retrovirus were transduced to CD45.1.2 WT or Mettl8−/− OT-I cells. 5 × 105 GFP+ cells were sorted 48 h after transduction and adoptively transferred into the tumor-bearing mice at 9 dpi. Mice were harvested at 19 dpi. (B) Tumor growth in each group of the mice in A. n = 6 per group. (C) Tumor growth in each group displayed in each replicate. n = 6 per group. (D) Tumor weight (left) and the absolute number of tumor infiltrating OT-I cells (right) from the mice in A. n = 6 per group. (E) Representative flow cytometry plots and cumulative data show the frequency of GFP in OT-I cells. n = 6 per group. (F) Representative flow cytometry plots (left) and cumulative data (right) show the frequency and absolute number of Tcf1+ Tim3− TPEX and Tim3+ Tcf1− TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (G) Representative flow cytometry plots (left) and cumulative data (right) show the frequency and absolute number of CX3CR1+ Tcf1− Int-TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (H) Representative flow cytometry plots (left) and cumulative data (right) show the frequency and absolute number of GzmB+, IFN-γ+, and perforin+ cells gated on tumor-infiltrating OT-I cells. n = 6 per group. Data are representative of two independent experiments. P value was calculated by two-way ANOVA (B) and two-tailed Student’s t test (D to H); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.