Extended Data Fig. 7: Glutathione synthesis modulates human NK cell effector functions through mitochondrial metabolism.

(a) Transcript expression of glutathione synthesis enzymes in human and mouse NK cells. (b) MFI of ThiolTracker Violet in SHIN1-treated human or mouse NK cells. (c) MFI of ThiolTracker Violet in Rosa26^cRNP or Phgdh^cRNP edited mouse NK cells cultured for 3 d in Ser+ or Ser- media with IL-15 after editing. (d) Viable cell yield of control edited or GCLC KO human NK cells 6 d post-electroporation. (e) Viable cell yield of control edited or GCLC^CRISPRa human NK cells 4 d post-electroporation. (f) MFI of pS6 in control edited or GCLC^CRISPRa human NK cells. (g) MFI of GzmB and perforin in K562-stimulated control edited or GCLC KO human NK cells. (h) Quantification of percent CD107a⁺ and CD107a MFI of degranulating cells of control edited or GCLC KO human NK cells stimulated for 4 h with K562 cells and IL-2/15. (i) MFI of pS6 in control edited or GCLC KO human NK cells. (j, k) OCR (j) and ECAR (k) of control edited or GCLC KO human NK cells. (l) Basal ECAR of control edited or GCLC KO human NK cells. (m) Basal, ATP linked, and maximal OCR of control edited or GCLC^CRISPRa human NK cells. (n, o) ECAR of control edited or GCLC^CRISPRa human NK cells. (p) MFI of MitoTracker Green FM in control edited or GCLC^CRISPRa human NK cells. Data represent mean ± SEM or individual paired donors across at least two independent experiments. Data are representative of (a, g, h, j-l, p) n = 6, (d) n = 8, (e) n = 16, (f) n = 11, (i) n = 8, and (m-o) n = 7 independent human donors, and (a) n = 3, (b) n = 6, and (c) n = 3 independent mice. *p < 0.05, **p < 0.01, ***p < 0.001 by (a, c, h, j, k, n) two-way ANOVA or (b, d-i, l, m, o, p) two-sided paired t-test. Exact p-values: (a) p = 0.0495 human, p = 0.0037, 0.0012, 0.0186 mouse; (b) p = 0.0417, 0.0015; (d) p = 0.0009; (e) p = (g) p = 0.0023, 0.0001; (h) p = 0.0025; (i) p = 0.0214; (l) p = 0.0184.