NLRP3 Inflammasome Activation Assay

Anti-Flag (A00187–200), anti-HA (A01244–100), anti-Actin (A00730–100), and protein G resin (L00209) were purchased from GenScript. Anti-NLRP3 (AG-20B-0014-C100) was purchased from Adipogen. Anti-GSDMD (ab209845) and anti-NEK7 (ab133514) were purchased from Abcam. Anti-ASC (67824) was purchased from CST. Anti-mouse caspase-1 was a kind gift from Dr Gabriel Núñez (University of Michigan Medical School). Ultra-pure LPS (tlrl-pb5lps), Nano-SiO2 (tlrl-sio), and poly(dA:dT)/lyovec (tlrl-patc) were purchased from InvivoGen. Nigericin (481990), doxycycline hydrochloride (10592–13–9), and EDTA-free protease inhibitor cocktail (11873580001) were purchased from Sigma. Salmonella strain SL1344 was originally from Dr Denise Monack (Stanford University). The constructs for NLRP3-Flag, NLRP3-SFP, and NEK7-HA have been previously described (17 (link)).