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. 2023 Apr 26;7(5):e0114. doi: 10.1097/HC9.0000000000000114

FIGURE 4.

FIGURE 4

Mincle and GSDMD promoted IL-1β secretion by small extracellular vesicles (sEVs). (A–E) Gsdmd −/− or Mincle −/− and heterozygous littermate mice (n=6) were exposed to Gao-binge ethanol feeding. (A and B) In all, 100 mg of liver explants was cultured overnight. sEVs were isolated from the cell culture media and analyzed by nanoparticle tracking with ZetaView for size distribution and total EV numbers/mL media. (C and D) Concentration of IL-1β in the collected sEVs was measured by ELISA. (E) IL-1β was immunoprecipitated from culture medium for western blotting with the indicated antibodies. Recombinant IL1-β (Rec) was included as a positive control. (F) EVs isolated from plasma of HC and patients with AH were analyzed by western blotting with antibodies IL1-β and the additional indicated proteins. Data were represent mean±SEM. ANOVA *p < 0.05, Abbreviations: AH, alcohol-associated hepatitis; EtOH, ethanol; GSDMD, gasdermin D; HC, healthy controls; K, knock-out; m, mature IL-1β; Mincle, macrophage-inducible C-type lectin; p, precursor; W, wild type.