Extended Data Fig. 5: Effector molecule expression of tumour-infiltrating REGNASE-1-null CD8⁺ T cells.

a, b, OT-I cells transduced with control sgRNA (mCherry⁺) or Regnase-1 sgRNA (ametrine⁺) were mixed at a 5:1 ratio and transferred into tumour-bearing mice (n = 5 mice), and tumour-infiltrating OT-I cells were analysed at day 7 for the expression of CD69, CD25, CD49a, KLRG1, ICOS, LAG3, PD-1 and CTLA4 (a, top) and CD44 and CD62L (b, top), and quantification of MFI of CD69, CD25, CD49a, KLRG1, ICOS, LAG3, PD-1 and CTLA4 (a, bottom) and frequency of CD44⁺CD62L⁻ cells (b, bottom). The numbers in graphs indicate the MFI (a, top). The numbers in plots indicate the frequency of CD44⁺CD62L⁻ cells (b, top). c–f, OT-I cells transduced with control sgRNA (mCherry⁺) or Regnase-1 sgRNA (ametrine⁺) were mixed at a 5:1 ratio and transferred into tumour-bearing mice, and analysed at day 7 (n = 10 mice) or day 14 (n = 10 mice). Flow cytometry analysis of expression of IFNγ (c, top), GZMB (c, bottom), TNF (e, top) and IL-2 (e, bottom) in TIL OT-I cells, and quantification of the numbers of IFNγ⁺ cells (d, top), GZMB⁺ cells (d, bottom), TNF⁺ cells (f, left) and IL-2⁺ cells (f, right) per gram of tumour (normalized to input). The numbers adjacent to outlined areas indicate the frequencies of IFNγ⁺ cells and the MFI of IFNγ in IFNγ⁺ cells (c, top), and the frequency of GZMB⁺ cells and the MFI of GZMB in GZMB⁺ cells (c, bottom), and the frequencies of TNF⁺ cells (e, top) or IL-2⁺ cells (e, bottom). Mean ± s.e.m. (a, b, d, f). *P < 0.05, **P < 0.01, ***P < 0.001. Two-tailed unpaired Student’s t-test (a, b) or two-tailed paired Student’s t-test (d, f). Data are representative of two (a–c, e) independent experiments, or pooled from two (d, f) independent experiments.

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