Fig. 7.
Impaired glycolysis and mTOR activity in preterm neonatal monocytes. a Effect of blocking glycolysis on cytokine responses to LPS stimulation (24 h) in mononuclear cells (2-sided paired t-tests; boxes and whiskers; 4 to 9 samples per condition; 2-DG = 2-deoxy-d-glucose); b Extracellular acidification rates (ECAR, normalized to total protein content), under baseline glucose-free conditions, after addition of glucose, oligomycin, and 2-DG (representative experiment from a 29 week gestation preterm sample); c Glycolytic capacity (cumulative data pooled from 4 independent experiments; 3–9 subjects per age group; mean ± SD; p value by Mann–Whitney U test); d Lactate production in mononuclear cells after LPS stimulation (24 h); p value = effect of age by 2-way ANOVA. Data from 9 to 10 subjects per age group; boxes and whiskers; e Effect of blocking glycolysis or translation (using cycloheximide, CHX) on the phagocytosis of C. albicans (Ca); boxes and whiskers; 3 subjects; Cyto-D = cytochalasin-D; f Depiction of signaling events between Toll-like receptor and Raf-1-mediated dectin-1 activation, mTOR phosphorylation, and increased glycolysis and protein synthesis; g Western blot image (cropped) of mTOR/4EBP1 expression and mTOR phosphorylation (monocytes; preterm sample from 29 weeks gestation; cropped images from same blot probed for mTOR, β-actin, and 4EBP1, and stripped/re-probed for phospho-mTOR); h Expression of mTOR-related genes (monocytes; 6–12 per age group; boxes and whiskers; 2-way ANOVA across age groups, only significant p values are shown)