FIGURE 2.

Dihydrotanshinone I specifically inhibits NLRP3 inflammasome activation. (A) LPS-primed BMDMs were treated with dihydrotanshinone I (10 μM) and then stimulated with ATP (45 min), nigericin (30 min), poly (I: C) (6 h), or SiO₂(6 h), or Pam3CSK4-primed BMDMs were treated with dihydrotanshinone I (10 μM) and stimulated with LPS (6 h). Western blot analysis of IL-1β (p17) and caspase-1 (p20) in supernatants (Sup.) and pro–IL-1β and pro–caspase-1 in whole lysates (Lys.) of BMDMs are shown in (A). (B) BMDMs were primed with LPS, exposed to DHT (10 μM), and then stimulated with nigericin, Poly (dA:dT) (6 h), and Salmonella typhimurium (6 h). Immunoblotting analysis of IL-1β (p17) and cleaved caspase-1 (p20) in culture supernatants (Sup.) and pro–IL-1β and pro–caspase-1 in whole lysates of BMDMs (Lys.). (C–E) Activity of caspase-1 (C), secretion of IL-1β (D), and production of TNF-α (E) in Sup. from samples described in (A). (F–H) Activity of caspase-1 (F), IL-1β (G), and TNF-α (H) in Sup. from indicated samples in (B). Coomassie blue staining was used as the supernatant loading control (A–B) and Lamin B as the lysate loading control (A–B). Data are represented as mean ± SEM from three biological replicates. Statistics were analyzed by multiple t-tests. *p < 0.05, **p < 0.01, ***p < 0.001, NS: not significant.