Figure 1.

Eri inhibits NLRP3 activation in vitro. (A) The structure of Eri. (B) BMDMs were treated with or without LPS (1 µg/ml) for 3 h, and then treated with nigericin (2 μM) and the indicated concentrations of Eri for 3 h. Mature IL-1β and p20 in supernatants or pro-IL-1β and pro-Casp-1 in lysates were determined by western blot. (C) BMDMs were treated with or without LPS (1 µg/ml) for 3 h, and then treated with nigericin (2 μM) and the indicated concentrations of Eri for 3 h. IL-1β, IL-18 and TNF-α protein levels were determined by ELISA. (D, E) Experiments were performed as described in (B, C), except that BMDMs were treated with Eri (5 nM) for indicated times. (F) BMDMs were treated LPS (1 µg/ml for 3 h), MSU (150 µg/ml for 1 h), nigericin (2 μM for 30 min) or ATP (2.5 mM for 30 min). Then, BMDMs were treated 5 nM Eri for 3 h, mature IL-1β and p20 in supernatants or pro-IL-1β and pro-Casp-1 in lysates were determined by western blot. (G) Experiments were performed as described in (F), except that IL-1β protein levels were determined by ELISA. All experiments were repeated at least three times. Bar graphs present means ± SD (**P < 0.01; *P < 0.05). n.s., not significant.