Supplementary Figure 3: PGE2 receptors EP1 and EP3 do not inhibit, but EP4 does inhibit, the NLRP3 inflammasome.
From: NLRP3 inflammasome inhibition is disrupted in a group of auto-inflammatory disease CAPS mutations
(a-i) BMDMs primed for 3.5 h with LPS. (a,b) The indicated concentration of EP1 agonist ONO-DI-004 or (c,d) EP3 agonist ONO-AE-248 was added 5 min prior to (a,c) nigericin (5 μM for 30 min) or (b,d) ATP (5 mM for 45 min). (e-g) The indicated concentration of EP4 agonists (e left,f) CAY10598 and (e right,g) ONO-AE1-32 were added 5 min before (e) nigericin (5 μM for 30 min) or (f,g) ATP (5 mM for 45 min). (h,i) An EP4 antagonist ONO-AE3-208 (2 μM) was added for 5 min prior to addition of ONO-AE1-329 (500 nM) for 5 min and then treated with (h) nigericin (5 μM for 30 min) or (i) ATP (5 mM for 45 min). (a-i) Total IL-1β release in cell supernatants analyzed by ELISA. (a-d,f,g,i) Immunoblot analysis of culture supernatants (SN) and lysates (LYS). *P < 0.005 (unpaired two-tailed t test). (a-i) Data are from one experiment representative of three separate experiments with similar results, (a-i) BMDM were derived from 2 mice per experiment mixed together, (a-i) three replicates, each replicate is one well + s.e.m.
