FIGURE 2.

β-GluCer promoted Mincle-dependent IL-1β production without triggering cell death in hepatic macrophages. Primary hepatic macrophages isolated from Mincle ^+/- and Mincle ^−/− mice were primed overnight with 10 ng/mL LPS and then stimulated with 20 µg/mL GluCer for 12 hours or 2.5 mM ATP for 1 hour. (A) Cell lysates were collected for western blot analysis of pro- and mature- (m) IL-1β, with β-actin used as a loading control. (B/C) Cell culture medium was collected and used to (B) measure for IL-1β concentration by ELISA or (C) measure LDH release to assess cytotoxicity. Data represent mean±SEM. Two-tailed unpaired Student t test. *p < 0.05, ***p < 0.001. Abbreviations: Mincle, macrophage-inducible C-type lectin; LPS, lipopolysaccharide.