WT (n = 14), Nlrp3–/– (n = 10), Asc–/– (n = 16), and Casp1–/– Casp11–/– (n = 9) mice were infected with 106
L. major promastigotes. Twenty-eight days later, the infected footpads were harvested and homogenized as detailed in the Methods. (A) Protein lysates from the representative footpads were run on an SDS-PAGE gel, and activation of caspase 1 was determined by Western blotting. GAPDH was used as a loading control. (B and C) IL-1β and IL-18 cytokine levels were determined by ELISA in the footpad lysates. Dunnett’s multiple comparisons test was performed to evaluate significance between multiple groups. Data were combined from 2 independent experiments and represent the mean ± SEM. ***P < 0.001; ****P < 0.0001.