Supplementary Figure 6: PKA inhibits the NLRP3 inflammasome in response to extracellular agonists but not particulate agonists.
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,c) The indicated concentration of 6-Bnz-cAMP or (b) 8-CPT-2-O-Me-cAMP was added 5 min before nigericin (5 μM for 30 min). (d) 6-Bnz-cAMP (50 μM) or 8-CPT-2-O-Me-cAMP (50 μM) was added 5 min prior to treatment with MSU (125 μg/ml for 4h) or silica (250 μg/ml for 5h). (e) Rp-cAMP was added 5 min before or (f) PKI was added 30 min before (e,f) 5 min treatment with db-cAMP (1 μM) or forskolin (50 μM) and followed by nigericin (5 μM for 30 min). (g) Rp-cAMP was added 5 min before or (h) PKI was added 30 min before (g,h) 5 min treatment with PGE2 (500 nM) followed by nigericin (5 μM for 30 min). (a,b,d-h) Total IL-1β release in cell supernatants analyzed by ELISA. (c-e) 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,b,d-h) three replicates, each replicate is one well + s.e.m.
