Extended Data Fig. 2: Gating strategy for the identification of T cell subsets in peripheral blood of melanoma patients.

Multiparametric fluorescence activated cell sorting analysis using the indicated gates. a Lymphocyte gate on side scatter/forward scatter; b single cell gate to exclude doublets; c live gating to exclude dead cells from subsequent gates; d CD3⁺ gate for T cells; e,f CD4⁺ and CD8+ gates for “helper” and “killer” T cell subsets, CD8 was detected with a PE-Cy7 labelled antibody for the T_reg panel (e) and with a FITC labelled antibody for the T maturation panel (f); g CD4⁺/CD25⁺/CD127^−/low regulatory T cells (T_reg); h naive (top left) and memory (bottom right) gates total T cells; i ETE (top) and CD31⁻ naive (bottom) gates for naive T cells; j naive (top left) and memory (bottom right) gates for CD8⁺ T cells; k CD8⁺ memory T cell subsets, the left bottom subset (CCR7⁻/CD27⁻) represents the T_IE cells.