Extended Data Figure 4: Characterization of the T-cell infiltrate in Braf^V600E/Pten^−/−/CAT-STA mice.

a, Distribution of T-cell subsets in Braf^V600E/Pten^−/− and Braf^V600E/Pten^−/−/CAT-STA tumours (n = 6). b, c, Representative flow cytometry plots to discriminate αβ-TCR T cells and γδ-TCR T cells (b), naive (CD62L⁺CD44⁻) and effector (CD62L⁻CD44⁺) T cells (pre-gated on CD3⁺CD8⁺ T cells), and one representative example of CD44/CD45RA staining (c). Quantification of naive (CD62L⁺CD44⁻CD45RA⁺), effector (CD62L⁻CD44⁺CD45RA⁻) and memory (CD62L⁺CD44⁺CD45RA⁻) T cells is indicated on the right (n = 6). d, Representative flow cytometry plots of FoxP3⁺ T regulatory cells (n = 6). e, Quantification and comparison of PD-1/Lag3 double-positive T cells in Braf^V600E/Pten^−/− and Braf^V600E/Pten^−/−/CAT-STA tumours (n = 12). f, Representative flow cytometry of PD-1- and Lag3-positive T cells (pre-gated on CD3⁺CD8⁺ T cells) in Braf^V600E/Pten^−/− tumours. g, Il2 transcripts present in sorted CD3⁺ T cells from Braf^V600E/Pten^−/− tumours and spleen (n = 10). h, Ifng transcripts present in sorted CD3⁺ T cells from Braf^V600E/Pten^−/− and Braf^V600E/Pten^−/−/CAT-STA mice (n = 10). i, Expression level of PD-L1 in whole tumour tissue from both mouse models assessed by qRT–PCR (n = 8). j, Flow cytometric analysis of PD-L1 expression of non-haematopoietic tumour cells (CD45⁻), CD45⁺CD11c⁺ dendritic cells (DC) and CD45⁺CD3⁺ T cells. Shown is a representative example as histogram (grey isotype, red Braf^V600E/Pten^−/−; blue, Braf^V600E/Pten^−/−/CAT-STA) with mean fluorescent intensity of n = 3 given each histogram (red, Braf^V600E/Pten^−/−; blue, Braf^V600E/Pten^−/−/CAT-STA). k, Percentage of Gr1⁺ cells within the CD11b⁺ fraction of the tumour immune cell infiltrate (n = 8; absolute numbers Braf^V600E/Pten^−/−: 1,047 ± 418 cells per gram tumour to Braf^V600E/Pten^−/−/CAT-STA: 739 ± 185 cells per gram tumour; P = 0.7429). All data are mean ± s.e.m., Mann–Whitney U test.*P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001; NS, not significant.