Fig. 4.

Glycolysis in neutrophils is inhibited and accompanied by immune dysfunction in LPS-tolerant model. To build the LPS-tolerant model, PMNs were treated with LPS (100 ng/mL) for 4 h or 8 h. a L-lactate concentrations in untreated PMNs and in those stimulated with LPS for 4 h and 8 h. b Apoptosis and c necrosis in untreated PMNs and in those stimulated with LPS for 4 h and 8 h. d Chemotaxis and e Phagocytosis in culture media of untreated PMNs and those stimulated with LPS for 4 h and 8 h. f Representative flow cytometry images of neutrophils stained with FITC and showing MFI in untreated PMNs and those stimulated with LPS for 4 h and 8 h. Neutrophils were seeded in Seahorse XFp analyzer culture plates (Seahorse Bioscience, North Billerica, MA, USA) and real-time OCR (g, i, j) and ECAR (h, k, l) were measured. ECAR reflected lactate production in untreated PMNs and those stimulated with LPS for 4 h and 8 h. One-way ANOVA with Brown-Forsythe and Welch ANOVA tests were performed. Error bars represent SD. Data are means ± SD of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001