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. 2021 Oct 14;12:750815. doi: 10.3389/fphar.2021.750815

FIGURE 1.

FIGURE 1

Dihydrotanshinone I (DHT) blocks NLRP3 inflammasome activation induced by nigericin and ATP in BMDMs. (A) Dihydrotanshinone I structure. (B) BMDMs were exposed to DHT (2.5–80 μM) for 24 h. (C–E) BMDMs were primed with LPS and then exposed to different concentrations (2.5, 5, or 10 μM) of dihydrotanshinone I, followed by the stimulation of nigericin for 0.5 h. Immunoblotting analysis of IL-1β (P17) and activated caspase-1 (P20) in cell supernatants (Sup.) are shown (C). Caspase-1 activity (D) and IL-1β (E) secretion were measured. (F–H) BMDMs were primed with LPS and then treated with different concentrations (2.5, 5, or 10μM) of dihydrotanshinone I, followed by the stimulation of ATP for 1 h. Western blot analysis of matured IL-1β (P17) and activated caspase-1 (P20) in culture supernatants (Sup.) are shown (F). Caspase-1 activity(G) and IL-1β (H) secretion are measured. (I and J) The secretion of TNF-a in the supernatant of cells treated as described in C (I) and F (J) were determined by ELISA. Coomassie blue staining was used as the loading control of the supernatant (C, F) and Lamin B was used as the lysate loading control (C, F). Results are represented as mean ± SEM from three biological replicates. One-way ANOVA was used to analyze the data. *p < 0.05, **p < 0.01, ***p < 0.001, NS: not significant.