Fig. 6
cMyc is required for splenic NK cell metabolic and functional responses. a, b Ex vivo splenic NK cells and T cells were isolated from CD45.2 GFP-Myc reporter mice or CD45.1 WT mice, mixed in a 1:1 ratio and activated for 18 h with IL-2/12 in the presence or absence of BCH. The ratio of fluorescence in CD45.2 and CD45.1 cells was calculated to give a measure of cMyc expression (CD45.2 GFP-Myc) adjusted for autofluorescence (CD45.1 WT). c–i Ex vivo NK cells were left unstimulated (US) or were activated for 18 h with IL-2/12 in the presence or absence of BCH. c Flow cytometry was used to measure the expression of CD71. d, e Analysis of NK cell extracellular acidification rate (ECAR) to assess basal glycolytic rate and glycolytic capacity. f, g Analysis of NK cell oxygen consumption rate (OCR) to assess rates of OXPHOS and maximal respiration. Flow cytometry was used to measure the expression of IFNγ (h) and granzyme B (i). Data are mean ± s.e.m. of 5 (b) or 6 (c, e, g-i) or representative of 5 (a) or 6 (d, f, h, i) mice. d–g Data were normalised to 200,000 cells. Statistical analysis was performed using a one-way ANOVA with Tukey post test (b, c, e, g, h, i); *p < 0.05, **p < 0.01. Oligo oligomycin, 2DG 2-deoxyglucose, Anti A, antimycin A, Rot rotenone, FCCP carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone
