Supplementary Figure 4: PGE2 receptors EP2 and EP4 inhibit the NLRP3 inflammasome, but EP4 mediates PGE2-induced inhibition.
From: NLRP3 inflammasome inhibition is disrupted in a group of auto-inflammatory disease CAPS mutations
(a-h) BMDMs primed for 3.5 h with LPS. (a,b) The indicated concentration of EP2 agonist ONO-AE1-259-01 was added 5 min before treatment with (a) nigericin (5 μM for 30 min) or (b) ATP (5 mM 45 min). (c) The EP2 antagonist AH6809 (2 μM) was added 5 min prior to addition of ONO-AE1-259-01 followed by ATP (5 mM 45 min). (d) CAY10598 (500 nM) was added 30 min after stimulation with nigericin and analyzed 50 min post initial nigericin stimulation. (e,f) EP2 agonist ONO-AE1-259-01 or (g,h) EP4 antagonist ONO-AE3-208 was added 5 min before treatment with PGE2 (500 nM) followed by (e,g) nigericin (5 μM for 30 min) or (f,h) ATP (5 mM 45 min). (a-h). Total IL-1β release in cell supernatants analyzed by ELISA. (b,c,f,h) Immunoblot analysis of culture supernatants (SN) and lysates (LYS). *P < 0.005 (unpaired two-tailed t test). (a-h) Data are from one experiment representative of three separate experiments with similar results, (a-h) BMDM were derived from 2 mice per experiment mixed together, (a-h) three replicates, each replicate is one well + s.e.m.
