Extended Data Fig. 2: CD8-MP and CD8-Naive CDR3α chain hydrophobicity and length analysis.

CD8-MP (CD8β⁺ CD44^hiCD122⁺) and CD8-Naive (CD8β⁺ CD44^loCD122^–) T cells were purified by FACS from the pooled spleen and lymph nodes of 9-week-old TCRβtg males and subjected to complete TCRα sequencing using the iRepertoire platform. N = 5 for CD8-MP and CD8-Naive samples. TCRα chains were assessed solely based on their predicted CDR3 segment, regardless of V-region usage. a, Histograms depicting grand average of hydropathy (GRAVY) values for CDR3 regions of the CD8-MP (red) and CD8-Naive (black) subsets for each mouse, n = 5 mice. Significance testing was performed with the paired, two-tailed Wilcoxon signed-rank test (W) and the paired, two-tailed Kolmolgorov-Smirnov test (K-S). Mouse 1 (CD8-MP CDR3 n = 11999, CD8-Naive CDR3 n = 29048), p = 0.2449 (W) and p = 1 (K-S); Mouse 2 (CD8-MP CDR3 n = 3327, CD8-Naive CDR3 n = 14736), p = 0.0511 (W) and p = 1 (K-S); Mouse 3 (CD8-MP CDR3 n = 2159, CD8-Naive CDR3 n = 26649), **p = 0.0017 (W) and p = 1 (K-S); Mouse 4 (CD8-MP CDR3 n = 2848, CD8-Naive CDR3 n = 24501), ***p < 0.0001 (W) and p = 1 (K-S); Mouse 5 (CD8-MP CDR3 n = 13524, CD8-Naive CDR3 n = 25367), **p = 0.0010 (W) and p = 1 (K-S). (n.s. not significant). b, Histograms depicting CDR3 lengths of the CD8-MP (red) and CD8-Naive (black) subsets for each mouse, n = 5 mice. Significance testing was performed with the paired, two-tailed Wilcoxon signed-rank test (W) and the paired, two-tailed Kolmolgorov-Smirnov test (K-S). Mouse 1 (CD8-MP CDR3 n = 11999, CD8-Naive CDR3 n = 29048), p = 0.2031 (W) and p = 1 (K-S); Mouse 2 (CD8-MP CDR3 n = 3327, CD8-Naive CDR3 n = 14736), p = 0.2031 (W) and p = 1 (K-S); Mouse 3 (CD8-MP CDR3 n = 2159, CD8-Naive CDR3 n = 26649), p = 0.4258 (W) and p = 1 (K-S); Mouse 4 (CD8-MP CDR3 n = 2848, CD8-Naive CDR3 n = 24501), p = 1 (W) and p = 1 (K-S); Mouse 5 (CD8-MP CDR3 n = 13524, CD8-Naive CDR3 n = 25367), p = 0.9102 (W) and p = 1 (K-S). (n.s. not significant).