BMDMs or THP-1 cells treated with 20 ng/mL PMA were plated in 6 well plates and 24 h later primed with 100 ng/mL LPS (Sigma) for 4 h before being treated to activate the inflammasome. Activation was by stimulated with Nigericin (20 µM) for 30 min; ATP (5 mM) for 30 min; silica (120 µg/mL) for 6 h, or MSU (200 µg/mL) for 6 h (InvivoGen). For Clostridioides difficile (C. difficile)-related inflammasome activation, cells were stimulated with freshly cultured C. difficile 630 (ATCC BAA1382) at a multiplicity of infection (MOI) of 100 for 6 h or primed with LPS (1000 ng/mL) for 2 h followed by 200 ng/mL of the microbe toxin B (Abcam) for 4 h.
