Figure 4.
Blockade of glycolysis partially inhibited TREM-1-mediated NLRP3 inflammasome activation in macrophages. (A) Macrophages (1×106 cells/well) were premixed with PBS control or 2-DG (5 mM) and then plated into 12-well plates with agonist anti-TREM-1 mAb (10 μg/mL). After 24 h, supernatants were analyzed for lactate production, n=3. (B, D) Macrophages were treated as in (A), and 6 h later, Tnf‐α and Pro-il-1β mRNA levels in macrophages were measured using qPCR, n=3. (C, E) Macrophages were treated as in (A). Twenty-four hours later, TNF‐α and IL-1β production in the supernatant was measured by ELISA, n=3. (F) NLRP3, pro-IL-1β, IL-1β p17, and caspase-1 p10 protein in cell lysate were detected by western blot, n=3. (G-J) Quantification of indicated protein levels in (F). n=3 biological replicates. Data are expressed as the mean ± SD. One-way ANOVA adjusted by Tukey's multiple comparison test was used. * P < 0.05, ** P < 0.01, and *** P < 0.001.