Fig. 3: Neutrophils adopt pentose cycle during oxidative burst.
a–c, Labelling patterns of ribulose-5-phosphate (a), DHAP (b) and G6P (c) from 1,2-13C-glucose tracing. Bar graphs represent mean ± s.d., n = 3 biological replicates (neutrophils isolated from different donors represented by individual dots). d, Metabolic flux distributions in stimulated neutrophils with or without oxidative burst. Numbers in blue next to the reaction indicate relative net flux, which are normalized to HK flux as 100. Values represent average results from multiple donors, with s.d. indicated in parentheses (n = 3 for no oxidative burst condition, n = 5 for oxidative burst condition). e, Schematic showing G6P labelling from 3-2H-glucose. HK activity generates 2H-labelled G6P, whereas pentose cycle generates unlabelled G6P. f, Labelling pattern for G6P from 3-2H-glucose. Bar graph represents mean value of n = 2 technical replicates indicated by individual dots. Labelling in PMA-stimulated oxidative burst was repeated in four independent experiments. g, PMA-induced oxidative burst is suppressed in TKT or TALDO1 KO HL-60 cells (mean ± s.d., n = 6 (WT) and n = 12 (TKT and TALDO1 KO) independent samples). h, Schematic illustrating expected average labelling enrichment from 1,2-13C-glucose when cells metabolize glucose: (1) 100% via glycolysis, (2) 100% via oxPPP with no recycling (no GPI flux) or (3) via full pentose cycle with reversed GPI net flux (that is, oxPPP flux is 300% of total glucose uptake). i, Average labelling enrichment of DHAP from 1,2-13C-glucose in human peripheral blood neutrophils stimulated with PMA (100 nM) with or without NOX inhibitor DPI (10 µM) or G6PDi (50 μM). Mean ± s.d., dots represent results in different biological replicates (neutrophils from different donors), n = 4 (PMA, PMA + DPI), n = 2 (PMA + G6PDi). j, Average labelling enrichment of DHAP from 1,2-13C-glucose from WT, TKT or TALDO1 KO HL-60 cells. Mean ± s.d. from independent samples measured in independent experiments; n = 2–5 in each specific condition, as indicated by individual dots. k, Average labelling enrichment of DHAP from 1,2-13C-glucose in human peripheral blood neutrophils stimulated ex vivo with zymosan A (300 μg ml–1), TNF-α (100 ng ml–1) or PsA (10:1 bacteria/neutrophils) with or without DPI (10 µM). Mean ± s.d., dots represent different biological replicates (different donors), n = 4 (zym, TNF-α), n = 2 (PsA).
