Fig. 2.

MIF regulates NLRP3-dependent release of IL-1 family cytokines. a Primary murine WT or Mif^−/− BMDMs were left untreated, primed with LPS alone (10 ng ml⁻¹), or primed with LPS before transfection of poly dA:dT (1 µg ml⁻¹) for 5 h. WT BMDMs were left untreated, primed with LPS (10 ng ml⁻¹), or primed with LPS before the addition of COR123625 (50 µM) for 2 h before transfection of b poly dA:dT (1 µg ml⁻¹) or c flagellin (250 ng ml⁻¹) for 5 h. Alternatively, following priming of WT BMDMs with LPS, cells were treated with d, g MSU (150 µg ml⁻¹), e, h silica (150 µg ml⁻¹), or (f, i) PBI-F2 peptide (100 µg ml⁻¹) for 6 h. Levels of a–f IL-1β and g–i IL-18 in cell culture supernatants were quantified by ELISA. j Primary WT or Mif^−/− BMDMs were primed with LPS (10 ng ml⁻¹) in the presence or absence of recombinant murine MIF (rMIF, 10 ng ml⁻¹) before stimulation with nigericin (5 µM) for 1 h. In addition, cells were treated with or without COR123625. Levels of IL-1β in cell culture supernatants were assessed by ELISA. k Primary WT or Mif^−/− BMDMs were primed with LPS (10 ng ml⁻¹) in the presence or absence of MIF-containing conditioned media from WT BMDM before stimulation with nigericin (5 µM) for 1 h. Levels of IL-1β in cell culture supernatants were assessed by ELISA. Data are expressed as means ± SEM, n = 3 mice. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001, one-way ANOVA with a correction provided by the Tukey's multiple comparisons test