Extended Data Fig. 8: Transient and stable expression of age-specific TCR.

a, Gating strategy transient transfection age-specific TCRs in HEK293T cells. b, Median MFI A2/M1₅₈ tetramer-PE staining of transiently expressed TCRs in HEK293T cells, dotted line indicates MFI threshold set by the parental cell line expressing no TCR (n = 3 independent experiments, median and IQR). c, Gating strategy and A2/M1₅₈ tetramer-PE staining with a normal (A2/M1₅₈-WT, lightly shaded), knockout CD8-binding site (A2/M1₅₈-KO open) and enhanced CD8-binding site (A2/M1₅₈-Enh, closed) tetramer of SKW-3-CD3⁺ and SKW-3-CD3⁺CD8⁺ TCR-expressing cell lines. d, Median MFI of A2/M1₅₈ tetramer-PE staining with a normal (A2/M1₅₈-WT, lightly shaded), knockout CD8-binding site (A2/M1₅₈-KO open) and enhanced CD8-binding site (A2/M1₅₈-Enh, closed) tetramer of SKW-3-CD3⁺ TCR-expressing cell lines (n = 2 independent experiments), dotted line indicates MFI threshold set by the parental cell line expressing no TCR. e, Gating strategy and CD69-PECy7 expression following peptide titration for public and age group specific TCRs (SKW-3-CD3⁺ cells). f, Percentage of maximum CD69-PECy7 MFI for age-specific TCRs expressed on SKW-3-CD3⁺ (open circles) and SKW-3-CD3⁺CD8⁺ (closed circles) cell lines following peptide titration, EC50 indicated by the dotted line (n = 3 independent experiments, median and range). Statistical analysis was performed using a two-sided Kruskal-Wallis with Dunn’s correction for multiple tests between the public and the age-specific TCRs (b) and a one-tailed Mann-Whitney U-test was performed for the EC50 between SKW-3-CD3⁺CD8^- SKW-3-CD3⁺CD8⁺ TCR-expressing cell lines (f). Exact significant p-values are indicated above the graphs.

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