Figure 2.
Mettl8 deficiency restricts tumor progression by promoting TPEXcell transition. (A) Schematic diagram of the adoptive transferred tumor model: CD45.1 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 2 × 106 CD45.2 WT or Mettl8−/− OT-I cells transfer at 9 dpi. Mice were harvested at 21 dpi. (B) Tumor growth in each group of the mice in A. n = 8 per group. (C) Survival curve in each group of the mice in A. n = 8 per group. (D) The absolute number of tumor infiltrating OT-I cells from the mice in A. n = 8 per group. (E and F) Representative flow cytometry plots and cumulative data show the frequency and absolute number of Tcf1+ Tim3− TPEX, Tim3+ Tcf1− TEX (E), and CX3CR1+ Tcf1− Int-TEX cells (F) gated on tumor-infiltrating OT-I cells. n = 6–8 per group. (G) 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 = 7 per group. (H) Representative flow cytometry plots (left) and cumulative data (right) show the frequency and MFI of GzmB, IFN-γ, and perforin gated on tumor-infiltrating Tcf1+ Tim3− TPEX, CX3CR1+ Tcf1− Int-TEX, and CX3CR1− Tcf1− TEX subsets. n = 5–6 per group. Data are representative of three independent experiments. P value was calculated by two-way ANOVA (B), Log-rank test (C), and two-tailed Student’s t test (D−H); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Refer to the image caption for details. Panel A shows schematic diagram of adoptive transfer tumor experiment timeline. Panel B shows line graph comparing tumor growth across experimental groups. Panel C shows Kaplan-Meier survival curve comparing survival of experimental groups. Panel D shows tumor infiltrating O T I cell numbers. Panel E shows flow cytometry plots and quantification of T c f 1 positive T P E X and T E X cells. Panel F shows flow cytometry plots and quantification of C X 3 C R 1 positive intermediate T E X cells. Panel G shows flow cytometry plots and quantification of G z m B, I F N gamma, and perforin expression. Panel H shows flow cytometry plots and M F I quantification across T cell subsets.

Mettl8 deficiency restricts tumor progression by promoting T PEX cell transition. (A) Schematic diagram of the adoptive transferred tumor model: CD45.1 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 2 × 106 CD45.2 WT or Mettl8−/− OT-I cells transfer at 9 dpi. Mice were harvested at 21 dpi. (B) Tumor growth in each group of the mice in A. n = 8 per group. (C) Survival curve in each group of the mice in A. n = 8 per group. (D) The absolute number of tumor infiltrating OT-I cells from the mice in A. n = 8 per group. (E and F) Representative flow cytometry plots and cumulative data show the frequency and absolute number of Tcf1+ Tim3− TPEX, Tim3+ Tcf1− TEX (E), and CX3CR1+ Tcf1− Int-TEX cells (F) gated on tumor-infiltrating OT-I cells. n = 6–8 per group. (G) 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 = 7 per group. (H) Representative flow cytometry plots (left) and cumulative data (right) show the frequency and MFI of GzmB, IFN-γ, and perforin gated on tumor-infiltrating Tcf1+ Tim3− TPEX, CX3CR1+ Tcf1− Int-TEX, and CX3CR1− Tcf1− TEX subsets. n = 5–6 per group. Data are representative of three independent experiments. P value was calculated by two-way ANOVA (B), Log-rank test (C), and two-tailed Student’s t test (D−H); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

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