Fig. 8

MIF interacts with NLRP3. a Immortalized BMDMs were left untreated, primed with LPS alone (100 ng ml⁻¹) for 5 h, or primed with LPS before inflammasome activation with nigericin (5 µM, 30 min). Lysates were immunoprecipitated with anti-MIF or anti-NLRP3 antibody, followed by western blot analysis with antibodies against NLRP3 and MIF. b WT BMDMs were primed with LPS alone (10 ng ml⁻¹) for 5 h, primed with LPS followed by NLRP3 activation with nigericin (5 µM) for 30 min, or primed with LPS followed by COR123625 (50 µM) treatment for 2 h prior to the addition of nigericin (5 µM) for 30 min. Levels of interaction between MIF and NLRP3 were assessed by FLIM-FRET microscopy. FLIM-FRET images presented are representative of three independent experiments. Scale bar = 50 μm. Graphs show changes in the amplitude-weighted average lifetime (τ Av Amp) of the donor (A488) due to proximity to the acceptor (A568). Combined data from three mice, analyzed by paired t test. *P < 0.05, **P < 0.01