Whole lungs collected from PBS-or HDMtreated wild type mice 1 day post-RV infection were subjected to Western blot (A and B) and qPCR analysis (C). (N= 7 from two different experiments, mean±SEM, *different from WT sham, †different from WT RV, p<0.05, two-way ANOVA with Tukey’s multiple comparisons test.) D. Differential immune cell counts in BAL of PBS-or HDM-sensitized and challenged, sham- or RV-treated wild type, NLRP3−/−, and IL-1β−/−mice (105 cells/ml) 2 days after treatment (N=7 from two different experiments, mean±SEM, *different from WT sham, †different from WT PBS RV or WT HDM RV, ‡ different from WT HDM sham, p<0.05, two-way ANOVA with Tukey’s multiple comparisons test). E and F. Airways responsiveness was measured in PBS-or HDM-sensitized and challenged, sham- or RV-treated wild type, NLRP3−/−, and IL-1β−/−mice 2 days after treatment. (N=4 from two different experiments, mean±SEM, *different from WT sham, †different from WT RV, p<0.05, two-way ANOVA). G. mRNA levels of type 2 cytokines in mice lungs. (N=7 from two different experiments, mean±SEM, *different from WT sham, †different from WT PBS RV or WT HDM RV, ‡ different from WT HDM sham, p<0.05 two-way ANOVA with Tukey’s multiple comparisons test). H and I. Recombinant IL-1RA (1 mg/kg body weight) was given after HDM treatment, one h before and 24 h after RV infection. BAL cell counts (H) and lung mRNA levels of type 2 cytokines (I) were measured 2 days post-RV infection. (N=7 from two different experiments, mean±SEM, *different from HDM-sham, †different from HDM RV, p<0.05, one-way ANOVA). For clarity, individual data points are not shown for panels B, F and G.