Extended Data Fig. 5: The antitumor effect of metformin is not mediated by conventional T cells.
From: Tumor hypoxia represses γδ T cell-mediated antitumor immunity against brain tumors
a,b, Survival of ctrl- and metformin-treated C57BL/6J mice depleted for CD8 T cells (a) or CD4 T cells (b) following injection with GL261 cells into the brain cortex. For T cell depletion, mice were injected intraperitoneally with antibody 1 and 2 days prior to tumor injection and every 7 days post-tumor injection. Data are representative of three independent experiments. Survival data were analyzed by log-rank test. c,d, Confocal microscopy of brain tumor tissue from Met-treated mice at 20 days post-tumor injection. Filled white arrows indicate perivascular γδ T cells. Empty white arrows indicate tumor-infiltrating γδ T cells (c). Cleaved Caspase-3 (Asp175) of tumor area with γδ T cells were shown (d). The images are representative of three independent experiments. Scale bar indicates 20 μm. e, Gating scheme of flow cytometry analysis for orthotopic HGG mice model. f-g, Flow cytometry quantification of CD4 T cells (f) and CD8 T cells (g) from ctrl- and metformin-treated C57BL/6 mice following 0 (n = 3), 10 (n = 5), 15 (n = 5) and 20 (n = 4) days after injection with GL261 cells into the brain cortex. Data are representative of four independent experiments. Error bars show mean ± SEM. Data were analyzed by a two-tailed unpaired Student’s t test. n.s., not significant (p > 0.05).
