Extended Data Fig. 1: Cytokine activation induces conserved and distinct metabolic changes in human and mouse NK cells.
From: Species-specific serine metabolism differentially controls natural killer cell functions
(a) Representative histograms and quantification of forward scatter and side scatter of freshly isolated naïve or 3 day IL-2/15-stimulated human NK cells. (b) Percent of 13C labelling of lactate, pyruvate, and citrate in naïve or 3 day IL-2/15-stimulated human NK cells cultured with [U-13C]-glucose for 24 h and analysed by GC/MS. (c) Representative histograms and quantification of tetramethylrhodamine ethyl ester (TMRE) MFI normalized to MitoTracker Green MFI in naïve or 3 day IL-2/15-stimulated human NK cells. (d) Representative histograms and quantification of MFI of phosphorylated AKTT308, phosphorylated S6S235/236, and phosphorylated AKTS473 of freshly isolated naïve or 3 day IL-2/15-stimulated human NK cells. (e) Quantification of MFI of CD69, KLRG1, and NKG2D of naïve or 3 day IL-2/15-stimulated human NK cells. (f) Schematic showing workflow of LC/MS analysis of NK cell metabolites from naïve or cytokine-stimulated cells. (g) Distribution across metabolic pathways of detected metabolites that were present in all samples. (h) Inertia barplot from PLS-DA analysis indicating number of components needed to capture inertia of sample dataset. (i) Variable Importance in Projection (VIP) scores of top 20 metabolites contributing to predictive model generated by PLS-DA analysis. (j) Heatmap showing log2 fold change of metabolites associated with serine/glycine metabolism pathways in naïve or stimulated NK cells compared to the average metabolite abundance of naïve cells. (k) Gating strategy for human NK cells. Data represent mean ± SEM or individual paired donors. Data are representative of (a, b) n = 4, (c) n = 6, (d) n = 5, (e) n = 7, and (j) n = 3 independent human donors and (j) n = 3 mice. *p < 0.05, **p < 0.01, ***p < 0.001 by (a) two-sided Student’s t-test, (b) two-way ANOVA, or (c-e) two-sided paired t-test. Exact p-values: (a) (left) p = 0.0001, (right) p = 0.0131; (b) p = 0.0045, 0.0089; (c) p = 0.0062; (d) p = 0.0251, 0.0250, 0.0109; (e) p < 0.0001, p = 0.0029, p = 0.0442. Created in BioRender. Li, J. (2025) https://BioRender.com/8kzb774.
