Panel A shows schematic diagram of M e t t l 8 detection in transfected cells. Panel B shows western blot showing effects of ginkgolic acid treatment. Panel C shows schematic diagram of ginkgolic acid treatment tumor model. Panel D shows line graph comparing tumor growth with ginkgolic acid treatment. Panel E shows individual replicate tumor growth curves for treatment groups. Panel F shows tumor weight in treated mice. Panel G shows infiltrating C D 44 positive C D 8 T cells. Panel H shows flow cytometry plots and quantification of t d T o m a t o and L y 108 expression. Panel I shows flow cytometry plots and quantification of C X 3 C R 1 positive intermediate T E X cells. Panel J shows flow cytometry plots and quantification of T P E X and T E X cells. Panel K shows flow cytometry plots and quantification of G z m B, I F N gamma, and Perforin. Panel L shows cumulative data of cytokine producing C D 8 T cell numbers. Panel M shows cumulative data of cytokine expression mean fluorescence intensity.
GA enhances the antitumor response of CD8 + T cells. (A) Schematic diagram of detecting Mettl8 expression in vitro: HEK293T cells were transfected with pMigR1-Mettl8-Flag plasmid, and Flag-tagged Mettl8 was detected by western blotting. (B) The detection included GA-treated HEK293T cells after transfection (top) or treated protein extracted from transfected HEK293T cells (bottom). (C) Schematic diagram of GA treatment to B16F10 tumor–bearing Mettl8-tdTomato-Flag mice: Mice were subcutaneously injected with 2 × 105 B16F10 cells, followed by GA treatment every 2 days from day 6 to day 12. Mice were harvested at day 13. (D) Tumor growth of the mice in C. n = 7 per group. (E) Tumor growth of the mice in C displayed in each replicate. (F and G) Tumor weight (F) and the absolute number of tumor-infiltrating CD44+ CD8+ T cells (G). n = 7 per group. (H) Representative flow cytometry plots (left) and cumulative data (right) show the MFI of tdTomato and Ly108 in tumor-infiltrating CD8+ T cells. n = 5–6 per group. (I and J) Representative flow cytometry plots and cumulative data show the frequency and absolute number of CX3CR1+ Tcf1− Int-TEX (I), Tcf1+ Tim3− TPEX, and Tim3+ Tcf1− TEX cells (J) gated on tumor-infiltrating CD44+ CD8+ T cells. n = 7 per group. (K) Representative flow cytometry plots (left) and cumulative data (right) show the frequency of GzmB, IFN-γ, and perforin gated on tumor-infiltrating CD44+ CD8+ T cells. n = 6–8 per group. (L and M) Cumulative data show the absolute number (L) and MFI (M) of IFN-γ, GzmB, and perforin gated on tumor-infiltrating CD44+ CD8+ T cells. n = 6–8 per group. Data are representative of three independent experiments. P value was calculated by two-way ANOVA (D) and two-tailed Student’s t test (F–M); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Source data are available for this figure: SourceData F6.