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. 2021 Mar 29;12:630380. doi: 10.3389/fimmu.2021.630380

Figure 1.

Figure 1

Inhibition of lactate dehydrogenase prevents NLRP3 inflammasome activation. (A) ELISA was used to determine the IL-1β levels in culture supernatants of mouse bone marrow-derived macrophages (BMDMs) that had been pretreated with 2DG or GSK2837808A for 1 h, followed by stimulation with nigericin for 45 min. (B) Determination by ELISA of IL-1β in the culture supernatant of THP-1-derived macrophages that had been pretreated with 2DG or GSK2837808A for 1 h, followed by stimulation with nigericin for 45 min. (C) Determination by ELISA of IL-1β in the culture supernatant of THP-1-derived macrophages that had been pretreated with GSK2837808A for 1 h, followed by stimulation with ATP, MSU, or alum for 4 h. (D) Immunoblot analysis of NLRP3 inflammasome molecules in culture supernatants (SN) and cell lysates (CL) of THP-1-derived macrophages that had been treated with or without 2DG or GSK2837808A for 1 h and then stimulated with nigericin for 45 min. The densitometric analysis of caspase-1 (p20) and IL-1β (p17) was shown in the right panels. (E) Immunoblot analysis of NLRP3 inflammasome molecules in cell supernatants and cell lysates of THP-1-derived macrophages treated with LDHA- or negative control- siRNA, and then stimulated with nigericin for 45 min. The densitometric analysis of caspase-1 (p20) and IL-1β (p17) was shown in the right panels. *P < 0.05; **P < 0.01. All results are presented as the mean ± SD of data from three independent experiments (n = 3 per group), and were analyzed with the ANOVA test followed by post hoc test.