Fig. 3: TRPV1 deletion suppresses NLRP3 inflammasome activation in microglia.

Primary mouse microglia from WT and TRPV1-KO pups were seeded in 96-well or 6-well plate overnight. Then cells were primed with100 ng/ml LPS for 3 h and subsequently challenged with 5 mM ATP for 30 min or 10 μM nigericin for 1 h to activate NLRP3 inflammasome. A–D IL-1β and TNF-α release in the supernatants were measured using ELISA method. E Cell lysates were immunoblotted with IL-1β p17 and Caspase-1 p10 antibody, and the results are summarized in (F, G). Cells were fixed, permeabilized and blocked, and then stained with anti-ASC antibody. H ASC speck formation was detected using confocal microscopy. (I) Statistics of the percentage of cells with ASC specks. Scale bars: 25 μm. Data are representative of three independent experiments. ***p < 0.001.