Extended Data Fig. 3: 1,2-13C glucose tracing and metabolic flux analysis indicate neutrophils switch to pentose cycle during oxidative burst induced by PMA (Related to Fig. 3).
a. Labeling pattern of selected PPP and glycolytic intermediates from 1,2-13C glucose in neutrophils stimulated with PMA (100 nM) for 30 min, with or without the treatment of DPI (10 µM). Mean ± SD, N = 3 biological replicates. Dots indicate results measured in neutrophils from each individual donor. *F6P peak may contain some glucose-1-phosphate, as these two compounds are not fully resolved on LCMS. b,c. Glycolysis and PPP reach pseudo-steady state at the peak of oxidative burst (~30 min post stimulation), as indicated by steady metabolite abundances (b) and labeling patterns (c) 25, 30, 35 min post- PMA stimulation. Bar graph show mean value of technical replicates when applicable. Trends have been repeated in two independent experiments. d. Relative net flux of key reactions determined in neutrophils isolated from different donors (indicated by different colors). Hatched bars are the fluxes in absence of oxidative burst (30 min after stimulation with 100 nM PMA, with 10 μM DPI treatment) and solid bars are the fluxes during oxidative burst (30 min after stimulation with 100 nM PMA). Error bars indicate the confidence intervals from each flux analysis.
