Panel A shows flow cytometry plots and quantification of C D 44 expression in O T I cells. Panel B shows flow cytometry plots and quantification of P D 1 expression in O T I cells. Panel C shows flow cytometry plots and quantification of Caspase and K i 67 expression in O T I cells. Panel D shows cumulative data of Caspase and K i 67 frequencies across T cell subsets. Panel E shows Cumulative data of G z m B, I F N gamma, and Perforin frequencies. Panel F shows schematic diagram of C R C liver metastasis experimental model. Panel G shows flow cytometry plots and quantification of T P E X to T E X ratio. Panel H shows flow cytometry plots and quantification of cytokine expression in C D 8 T cells. Panel I shows schematic diagram of melanoma lung metastasis model and survival. Panel J shows schematic diagram of adoptive transfer infection experimental timeline. Panel K shows absolute number of P 14 cells. Panel L shows flow cytometry plots and quantification of T cell subsets.
Mettl8 promotes T PEX differentiation without affecting their proliferation and apoptosis. (A and B) Representative flow cytometry plots and cumulative data show the frequency of CD44 (A) and PD-1 (B) OT-I cells infiltrating in tumors. (C) Representative flow cytometry plots and cumulative data show the frequency of caspase and Ki67 in tumor-infiltrating OT-I cells. (D) Cumulative data show the frequency of caspase and Ki67 in Tcf1+ Tim3− TPEX, CX3CR1+ Tcf1− Int-TEX, and CX3CR1− Tcf1− TEX subsets. (E) Cumulative data show the frequency of GzmB, IFN-γ, and perforin in tumor-infiltrating OT-I subsets mentioned above. (F) Schematic diagram of the classic CRC liver metastases model: Mettl8fl/flCd4cre and Mettl8fl/fl mice were intrasplenically injected with 2 × 105 MC38 cells (left), and imaging of livers on day 21 after injection (right). (G) Representative flow cytometry plots and cumulative data show the ratio of Tcf1+ Tim3− TPEX to Tim3+ Tcf1− TEX cells gated on CD44hi CD62Llo CD8+ T cells of the livers from mice in F. (H) Representative flow cytometry plots and cumulative data show the frequency of GzmB, IFN-γ, and TNF-α gated on CD44hi CD62Llo CD8+ T cells from the livers of mice in F. (I) Schematic diagram of the classic melanoma lung metastases model: Mettl8fl/flGzmbcre and Mettl8fl/fl mice were i.v. injected with 2 × 105 B16F10 cells (top) and the survival curve (bottom). (J) Schematic diagram of adoptive transfer model: CD45.1.2+Mettl8−/− or WT P14 CD8+ T cells were adoptively transferred into CD45.2+ WT recipients, followed by LCMV-clone 13 (LCMV-C13) infection 24 h later and then analyzed on 30 dpi. (K) Statistical analysis show the absolute number of P14 cells from the spleens of mice in J. (L) Representative flow cytometry plots and cumulative data show Tcf1+ Tim3− TPEX, Tim3+ Tcf1− TEX, and CX3CR1+ Tcf1− Int-TEX cells gated on P14 cells from the spleens of mice in J. n = 4–8 mice per group. Data are representative of two independent experiments. P value was calculated by two-tailed Student’s t test (A–H, K, and L) or Log-rank test (I); *P < 0.05; **P < 0.01; ***P < 0.001.