(A) Total cell lysates (TCL) from COPTG transgenic (CARD16, CARD17, CARD18) and wild-type (WT) BMDM were analyzed for GFP-COP expression by immunoblot and immunoprecipitation (IP).
(B and C) WT, COPTG, and Casp1−/− BMDM were primed with Pam3CSK4 (Pam) (1 μg mL−1, 4 h) and (B) treated with MSU crystals (180 μg mL−1, 6 h), silica (200 μg mL−1, 6 h), CPP (200 μg mL−1, 6 h), ATP (5 mM, 30 min), or nigericin (Nig) (5 μM, 45 min), or (C) transfected with p(dA:dT) (1 μg mL−1, 6 h), LeTx (0.5 μg mL−1, 6 h), or flagellin (0.5 μg mL−1, 6 h), and supernatants (SN) were analyzed for secreted IL-1β, IL-18, and TNF by ELISA (n = 3, mean ± SD). *p < 0.05. The dotted line indicates that Casp1−/− BMDM are only present in the primed and activated treatment group. Untreated and primed control cells are the same for different treatment groups.