Extended Data Fig. 2. LC reverses the LA-suppressive effects in CD8+ Teff cells.

a, Volcano plot of differential expression of CD8⁺ Teff cells treated with or without 10 mM LA for 3 hrs. CD8⁺ Teff cell activation relative genes were annotated. n = 3. b, Signature genes expressed by CD8⁺ Teff cells treated with LA and/or LC, identified through RNA-sequencing (RNA-seq) analysis. n = 3. c, IL-2 and IFN-γ in supernatant, detected by ELISA, of CD8⁺ Teff cells treated with 10 mM LA and/or LC/LiCl/Na₂CO₃/NaCl for 24 hrs. n = 4. d, CD25 expression (MFI), analyzed by flow cytometry, of CD8⁺ Teff cells treated as in c. n = 4. e, Proliferation, analyzed by flow cytometry, and cell counts of CTV-labeled CD8⁺ T cells treated with 10 mM LA and/or LC/LiCl/Na₂CO₃/NaCl for 72 hrs. n = 8. f, OVA-peptide primed OT-1 mice derived CD8⁺ Teff cells were treated with 10 mM LA plus LC, LiCl, Na₂CO₃ or NaCl for 24 hrs, and incubated with OVA-B16 tumor cells for 8 hrs. CD45^- cell apoptosis was determined by flow cytometry. n = 4. g, Relative abundance of mitochondrial M+3 lactate in CD8⁺ Teff cells pretreated with LC and/or DCA or Oxamate under the 11.1 mM ¹³C-glucose culture (n = 4). IL-2 (n = 7) and IFN-γ (n = 5) in supernatant, CD25 expression (n = 4), of CD8⁺ Teff cells treated with LC and/or DCA/Oxamate under the 11.1 mM glucose culture. h, Relative abundance of mitochondrial M+3 lactate in CD8⁺ Teff cells pretreated with LC and treated with or without 10 mM ¹³C-lactate under the 2 mM ¹³C-glucose culture (n = 4). IL-2 (n = 5) and IFN-γ (n = 5) in supernatant, CD25 expression (n = 5), of CD8⁺ Teff cells treated with LC and/or LA under the 2 mM glucose culture. Data are mean±s.e.m. n = biological replicates unless stated otherwise. P values were calculated using one-way ANOVA for Dunnett’s multiple comparisons test (c, d, f, g and h), ns (not significant).

Source data