Figure 2. Fenamate NSAIDs inhibit ASC speck formation and caspase-1 activation.

(a) iBMDMs stably expressing ASC protein conjugated to mCherry were primed with LPS (1 μg ml⁻¹, 2 h) then pre-treated with selected drug (100 μM, 15 min) before stimulation with ATP (5 mM, 30 min) under live microscopy. Formation of ASC specks (examples indicated by white arrows) was quantified (lower right) and presented as mean % specks counted versus vehicle+s.e.m (n=4). Scale bars are 20 μm. (b,c) THP-1 cells were primed with LPS (1 μg ml⁻¹, 4 h) and pre-treated with NSAID (200 μM, 15 min) before stimulating with nigericin (10 μM, 1 h). Supernatants were taken and analysed for IL-1β by ELISA (b) and the p10 active subunit of caspase-1 by western blot (c). ELISA data are presented as mean % IL-1β release versus vehicle (DMSO) control+s.e.m (n=3). *P<0.05, ***P<0.001 determined by one-sample t-test versus hypothetical value of 100%.