Inflammasome Protein Analysis by Western Blot

Treated cells were washed, detached with scraping, pelleted, resuspended with Nupage SDS loading buffer (ThermoFisher Sci.), and lysed with three 10s pulses at 40% sonication power (Heat Systems Ultrasonics, Plainview, NY). Lysates were loaded into NuPage 12% or 4–12% Bis-Tris gel wells (ThermoFisher Sci.) for gel electrophoresis, then transferred to nitrocellulose membrane with iBlot and developed using Inflammasome Antibody Sampler Kit #32961T (CST Danvers, MA) or rabbit anti-LTA or LT-B as previously described [46 (link)]. Blots were imaged with Pierce^™ ECL Western Blotting Substrate (ThermoFisher Sci.) and Amersham Imager 600 and quantified using ImageQuant LT (GE Healthcare, Pittsburgh, PA).