Extended Data Fig. 3 |. Cytosolic LPS-induced IL-18 release from murine cells is NLRP3-dependent.

a, b, c, WT or NLRP3-deficient iBMDMs were primed with extracellular LPS, or left unprimed, and electroporated with LPS (or PBS) in the presence of absence of MCC950. LDH, IL-18 and IL-1β release into supernatant was quantified after 2 h. n=3 biological replicates. d, e, LPS-primed WT iBMDMs were electroporated with LPS (or PBS) in the presence or absence of MCC950. IL-18 and IL-1β were immunoprecipitated from supernatants and analyzed by immunoblot (representative of three biological replicates). f, Immunoblot analysis caspase-4-deficient THP1 cells reconstituted with caspase-11 by retroviral transduction (representative of two independent repeats). g, Caspase-4-deficient THP1 macrophages expressing caspase-11 were primed with extracellular LPS, or left unprimed, and electroporated with LPS (or PBS) in the presence of absence of MCC950. LDH, and IL-18 release into supernatant was quantified. n=6 biological replicates. Bars and error bars represent mean ± SEM. Statistical significance was determined by two-way ANOVA (a,b,c) or one-way ANOVA (g) with Tukey’s multiple comparisons test: ns = not significant (p > 0.05). For gel source data, see Supplementary Figure 1.