Fig. 6

BRISC deficiency suppresses alcohol-induced NLRP3 inflammasome activation in liver. (A-E) WT and Abro1^−/− or Brcc3^−/− mice were subjected to chronic-plus-binge ethanol feeding for 10 days. Serum LPS levels were analyzed using ELISA (A). Hepatic NLRP3 and IL-1β expression was evaluated by real-time PCR (B, C). Western blot analysis of the indicated proteins in mice liver (D, E). (F) WT and Abro1^−/− KCs were primed with 500 ng/ml LPS for 4 h, then stimulated with the conditioned media from in vitro cultured primary hepatocytes of ethanol diet-fed WT mice for 30 min. The IL-1β level in the culture supernatants was measured. (G) K63-linked ubiquitination of NLRP3 in KCs isolated from mice livers fed with ethanol or control diet. The KCs from each group were pooled from three mice. Data are presented as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001; two-tailed unpaired t-test.