Extended Data Fig. 1: TCR stimulation strength controls the magnitude of T cell activation during priming.

a, Amino acid sequences of indicated Gp33 variants with alterations highlighted⁷⁵. b,c, Percentages of GFP (Nr4a)⁺ P14s in vitro stimulated by indicated concentrations of Gp33 peptides (b) or MC38 tumor cells stably expressing indicated Gp33 variants (c). d, LLC-Gp33 tumor growth in C57BL/6 mice, n = 7–12. e, Percentages of mCherry⁺ LLC-Gp33 at D18 post tumor inoculation, n = 5 per group. f, LLC-Gp33 tumor growth in Rag-KO mice, n(M9) = 5, n(C6M9) = 6. g, Representative flow plot of P14 percentages in tdLN, related to Fig. 1b. h,i, Percentages of CD44⁺(h, n = 6–8) and PD1⁺ (i, n = 3–8) P14s in tdLN. j,m, GMFI of GFP, n = 5. Gated on total (j) or CD44⁺ (m) P14-Nur77-GFP CD8⁺ T cells. P value 0.0022 (j). k, GMFI of Tox (left) and Tpex percentages (right) in Rag-KO, n = 4–6. Gated on CD44⁺ tdLN-P14s. P value 0.0029 (left). l, Percentages of CD62L⁺ P14s in tdLN, n = 4–6. Gated on CD44⁺ P14s. n, Percentages of PD1⁺Tim3⁺ P14s in tdLN, n = 5–9. d–f,j,k,m, Representative data of two independent experiments, two-tailed unpaired Student’s t test (mean ± s.e.m.), P value ** < 0.01, **** < 0.0001, ns represents not significant. h,i,l,n, Representative data of more than three independent experiments, n represents numbers of mice per group, Multiple two-tailed unpaired t test (mean ± s.e.m.), FDR = 0.05, q value indicated in each graph.

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