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. 2023 Feb 1;26(3):106062. doi: 10.1016/j.isci.2023.106062

Figure 1.

Figure 1

Tivantinib inhibits nigericin-induced IL-1β release and pyroptosis

(A and B) ELISA analysis of IL-1β and IL-18 in supernatants (SN) of LPS-primed BMDMs treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 30 min.

(C–F) Western blot analysis of IL-1β and caspase-1 (p20) in SN and pro-IL-1β, pro-caspase-1 in cell lysates (Input) of LPS-primed BMDMs treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 30 min. (D, E) ELISA analysis of IL-6 (D) and TNF-α (E) in SN of LPS-primed BMDMs treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 30 min. (F) LDH release in SN of LPS-primed BMDMs treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 30 min.

(G) Western blot analysis of Gsdmd cleavage in Input of Flag-Gsdmd reconstituted Gsdmd−/− iBMDMs cells primed with LPS for 3 h in the presence of various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 1 h.

(H) ELISA analysis of IL-1β in SN of PMA-differentiated THP-1 cells treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 1 h.

(I) Western blot analysis of IL-1β and p20 in SN and pro-IL-1β, pro-caspase-1 in Input of PMA-differentiated THP-1 cells treated with various concentrations of Tivantinib (as labeled) and then stimulated with 5 μM nigericin for 1 h. Data represent means ± SEM from four biological duplicates (A, B, D-F, and H). Statistical analysis was performed using one-way ANOVA of means test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.