Extended Data Fig. 1: Lactic acid upregulates TXNIP and ARRDC4 levels in CD8⁺ T cells and Treg cells, related to Fig. 1.

(a) The table of all differentially expressed genes of RNA-seq in both CD8⁺ T cells and Treg cells between the control and lactic acid-treated groups. (b, c) Volcano plot of CD8⁺ T cells (b) and Treg cells (c) treated with or without 10 mM lactic acid. (d, e) QPCR (d, n = 3 independent samples) and western blotting (e) analysis of human CD8⁺ T cells activated by anti-CD3/CD28 antibodies and treated with vehicle control, 10 mM lactic acid or 10 mM sodium lactate. (f-h) Flow cytometric and quantitative analysis of TNF (f), IFN-γ, GZMB (g) and IL-2 (h) expression of activated mouse CD8⁺ T cells treated with vehicle control, 10 mM lactic acid or 10 mM sodium lactate. (i) QPCR analysis of function-associated gene expression in human CD8⁺ T cells treated with or without 10 mM lactic acid (n = 3 independent samples). (j) QPCR analysis of function-associated gene expression in human iTreg cells treated with or without 10 mM lactic acid (n = 3 independent samples). (k) Flow cytometric and quantitative analysis of Treg suppressive capacity using in vitro suppression assays. Treg cells and CTV-labelled mouse CD8⁺ T cells were co-cultured for 48 h and treated with or without 5 mM lactic acid for 24 h (control: n = 4 cultures, lactic acid: n = 3 cultures). The data were representative of three independent experiments and presented as mean ± s.e.m. Statistical significance was calculated by the unpaired two­tailed Student’s t­test (d and f-h) and two-way ANOVA followed by Sidak’s multiple comparisons test (i-k). ns, not significant.

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