Extended Data Fig. 1: Flow cytometry analysis of naïve T cells from young and aged WT mice.
(a) Representative gating strategy used for FACS staining of T cells. (b) Numbers (#) of CD8+ T cells and (c) numbers (#, left) and frequency (%, right) of CD4+ T cells in various tissues of Y-WT and A-WT mice. (d) Representative dot-plots of IFNγ and TNFα expression (intracellular FACS staining) in LN CD4+ and CD8+ T cells and (e) summary results (frequency, left; and numbers, right) of IFNγ+, GzmB+, IL10+, IL17+, TNFα+ CD4+ and CD8+ cells in indicated tissues of Y-WT and A-WT mice. (f) Heatmap of surface proteins expression in FACS-stained CD4+ and CD8+ T cells using a modified LEGENDScreen™ Mouse PE Kit. (g) Gating strategy used for quantification of exhaustion-associated markers in CD8+ T cells. (h) Frequency of CD8+ T cells expressing exhaustion markers in PeC; and (i and j) representative dot plots for expression of Tox1 in indicated subpopulation of LN CD8+ cells with or without expression of Ly108 and CD69 (i), and CD101 and PD1 (j). In b, c, e and h, shown is mean number or frequency ± SEM, where each symbol is for a single mouse, n = 5-10 (b, c), n = 5 (e), and n = 5-6 (h) mice per group. Experiments were reproduced three times. * P < 0.05**; P < 0.01; *** P < 0.001 in unpaired Mann-Whitney test.
