Supplementary Figure 1: PGE2 does not induce degradation of NLRP3 inflammasome components but rapidly inhibits NLRP3 activation in macrophages.
From: NLRP3 inflammasome inhibition is disrupted in a group of auto-inflammatory disease CAPS mutations
(a-d) LPS-primed BMDM. (a) Lysates (LYS) from PGE2 (500 nM) treatment for 30 min. (b) The indicated concentration of PGE2 was added for 5 min followed by stimulation with (left) nigericin (5 μM for 30 min) or (right) ATP (5 mM for 45 min). (c) PGE2 (500 nM) was added 30 min after stimulation with nigericin and analyzed 50 min post initial nigericin stimulation. (d) 16,16-dimethyl PGE2 (16-DM, 500 nM) or 15-keto PGE2 (15-keto, 500 nM) were added 5 min prior to nigericin or ATP stimulation. (e-g) PMA-differentiated THP-1 cells were treated with (e,f) the indicated concentration of PGE2 for 5 min prior to stimulation with (e) nigericin (5 μM) and LPS (50 ng/ml) for 30 min or (f) ATP (5 mM) and LPS (50 ng/ml) for 45 min. (g) 16,16-dimethyl PGE2 (16-DM, 250 nM), or 15-keto PGE2 (15-keto, 250 nM) were added 5 min prior to nigericin. (b-g) Total IL-1β release in cell supernatants analyzed by ELISA. *P < 0.005 (unpaired two-tailed t test). (a-g) Data are from one experiment representative of three separate experiments with similar results, (a-d) BMDM were derived from 2 mice per experiment mixed together, (b-g) three replicates, each replicate is one well + s.e.m.
