Fig. 1: LPS increases glycolysis as well as lactate production by BM neutrophils.
a Flow cytometry quantitative analysis of 2-NBDG-glucose uptake by BM neutrophils (CD11bhighLy6Ghigh cells; n = 6) 4 h following i.p. administration of LPS in vivo in wild-type (WT) mice. b Gene expression of glycolytic enzymes in sorted BM neutrophils from WT mice following LPS treatment (n = 3, PBS; n = 5, LPS). On each box, the bottom, middle, and the top edges indicate the 25th, 50th, and 75th percentiles, respectively. The whiskers extend to the most extreme data points. c Quantitative analysis and representative histogram plot showing mean fluorescent intensity (MFI) of ROS production in BM neutrophils following LPS administration (n = 9). d Percentage of HIF-1α+ neutrophils in the BM following LPS administration (n = 11). e Quantitative analysis and representative histogram plot of LDHA expression in BM neutrophils following LPS treatment (n = 7). ***p(0.0003). f BM lactate levels in WT mice treated with PBS, LPS, or LPS followed by α-Ly6G Ab (n = 7). g Lactate levels released from isolated BM neutrophils treated ex vivo with PBS or LPS (120 ng/ml; n = 4 mice).**p(0.0063). h MCT4, MCT1, and GPR81 (yellow) distribution on BM CD11b+ (green)/Ly6G+ (red) neutrophils visualized and quantified by ImageStream analysis. Images are from one representative experiment out of three. Scale bar indicates 7 μm. i Quantitative analysis of MCT4 expression on BM neutrophils 4 h following LPS administration (n = 7). j A scheme of the proposed mode of action of LPS in lactate production by BM neutrophils. Data are represented as mean ± SEM from 3 to 4 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, Student’s two-tailed unpaired t test (a, c–e, g, i), one-way ANOVA with Tukey’s post hoc test (f, h) or two-way ANOVA with Tukey’s post hoc test (b). See also Supplementary Fig. 1.
