Extended Data Fig. 4: Treated CD8+ T cells maintain a heritable functional memory-like program.
(a) Representative FACS plots showing the gating strategy for sorting dysfunctional human CD8+ T cells on day 14 after chronic TCR plus TGFβ1 stimulation, and day 28 expression of IFNG and CD103 in PMA/ionomycin-stimulated CD8+ T cells after treatment. (b) Summary bar graphs showing frequency of IFNG + TNFα + CD8+ T cells, and (c) MFI of CD101 in sorted CD8+ T cells after PMA/ionomycin stimulation on day 28. (d) Representative FACS plots showing CD8+ T cell divisions and expression levels of IL-7R in CFSE-labeled CD8+ T cells from each condition as described in Fig. 4a on day 35 (after 7 day-rest under homeostatic conditions). (e) MFI of IL-7R in divided CD8+ T cells on day 35. (f) Representative FACS plots showing CD8+ T cell divisions and expression levels of CD103 in CFSE-labeled CD8+ T cells on day 35. (g) MFI of CD103 in divided CD8+ T cells on day 35. (h) Bar graphs showing frequencies of divided CD8+ T cells (≥3 proliferation cycles), and (i) MFI of IL-7R and (j) CD103 in CD8+ T cells on day 35. (k) Bar graphs showing MFI of TNFα and (l) Perforin in IFNG + CD8+ T cells after PMA/Ionomycin stimulation on day 35. N = 3 (panels a-c) or 4 (panels d-l) biological replicates per group, representative of 2–3 independent experiments. Adjusted P-value ** p < 0.01, *** p < 0.001, **** p < 0.0001 comparisons were made using one-way ANOVA with Tukey’s multiple comparisons as indicated (panels b, c, h–l) or one-way ANOVA with Bonferroni’s multiple comparisons relative to Dysf. group (panels e, g). Error bars = mean ± SEM.
