Figure 1.

Nlrp3 dampens pro-inflammatory cytokine response. (A) Wild type, Aim2^−/− and Nlrp3^−/− macrophages were infected with Francisella tularensis (Ft) live vaccine strain (LVS) at an multiplicity of infection (MOI) of 100. The cell lysates were prepared 24h post-infection and western blot analysis was performed to determine levels of bioactive caspase1 and IL-1β. The western blot images are representative of three independent experiments. β-actin was used as a loading control. The uncropped version of (A) is shown in Supplementary Figure S1. (B) The quantification of the bands (n=3 blots) is shown and expressed as relative intensity units (RIU). (C,D) The wild-type and the Nlrp3^−/− macrophages were infected with the indicated MOI of Ft LVS and treated with gentamicin to kill extracellular and adherent bacteria after 2h of infection and the infection with intracellular bacteria was allowed to proceed for 22h. The IL-1β (C) and TNF-α (D) levels were measured in the culture supernatants after 24h post-infection. The results are representative of three independent experiments. The data in B–D were analyzed by ANOVA with the Tukey-Kramer test, and a p-value of 0.05 or less was considered significant. ^*p<0.05; ^**p<0.01; ^***p<0.001.