Figure 1.

The LRRs of NLRP3 are dispensable for inflammasome activation. (A) Schematic representation of the LRR domain substitution construct. (B) Representative western blot of FLAG-tagged NLRs from HEK293T cell lysate. 10 μg total protein was loaded in each well. (C,D) IL-1β response of HEK293T cells expressing ASC, pro-Caspase-1, pro-IL-1β, and NLRP3, NLRP2, or the LRR domain substitution chimera NLRP332. (C) At 4-h post-transfection, cells were infected with Fn U112 (MOI 100). (D) 18–20 h post-transfection, cells were infected with Listeria monocytogenes (Lm) (MOI 20; 6 h) or treated with 100 μM H₂O₂ (1 h) or 5 μM nigericin (2 h). IL-1β in culture supernatant was measured by ELISA. Data represent means ± SEM for a minimum of three independent experiments. ^*p < 0.0001 for comparison with respective untreated controls; ^#p < 0.0001, two-way ANOVA followed by Tukey (^*) and Sidak's (^#) multiple comparison test.