Fig. 4. Fc–IL-4 augments glycolytic metabolism of CD8⁺ T_TE cells through STAT6 signalling and PI3K–AKT–mTOR axis.

a,b, Real-time ECAR analysis (a) and average basal glycolysis, glycolytic capacity and reserve (b) of ex vivo-induced CD8⁺ T_TE cells (n = 5 biological replicates). mpH, milli-pH. c, Volcano plot of altered metabolites in ex vivo-induced CD8⁺ T_TE cells treated with Fc–IL-4 (n = 4 biological replicates) versus PBS (n = 3 biological replicates). d–f, Experimental setting as described in Fig. 1g. Shown are unsupervised UMAP clustering of PMEL CD8⁺ TILs based on the 1,667 genes involved in KEGG-defining metabolic pathways (d), cell proportion in each cluster (e) and systematic expression comparison of carbohydrate metabolisms among top four clusters (f). g,h, T cell counts (g) and frequencies of granzyme B⁺IFNγ⁺ (h) among ex vivo-induced CD8⁺ T_TE cells with or without 2-DG (n = 4 biological replicates). i,j, Schematic illustration of single-cell ATAC and gene coprofiling of IL-4 versus PBS-treated ex vivo-induced CD8⁺ T_TE cells and a joint ATAC–gene UMAP (i), and volcano plot showing differentially active motifs (j). k,l, Experimental setting as described in Fig. 1g. Shown are signalling pathways regulated by DEGs (k) and top 20 ranked upstream regulators predicted from DEGs (l) in PMEL CD8⁺ TILs. m, Western blot analysis of indicated proteins in ex vivo-induced CD8⁺ T_TE cells (n = 3 biological replicates). n–p, Relative basal glycolysis (n) (n = 5 biological replicates), T cell counts (o) and granzyme B MFI (p) (n = 3 biological replicates) in Fc–IL-4-treated ex vivo-induced OT1 and OT1^STAT6-KO CD8⁺ T_TE cells (normalized by that in the PBS group) with or without indicated inhibitors. All data represent mean ± s.e.m. and are analysed by two-sided unpaired Student’s t-test (b and g,h), two-tailed Mann–Whitney test (j), right-tailed Fisher’s exact test (k) or one-way ANOVA and Tukey’s test (n–p). Schematics in i created using BioRender (https://Biorender.com).

Source Data