Figure 4.

NLRP3 deficiency reduces number of neutrophils and amount of NETs in lungs of pristane-induced DAH female, but not in male mice (A–C) Left column shows results with male mice (♂), right with female mice (♀). (A) Flow-cytometry analysis of BAL from pristane-induced DAH WT and NLRP3 -/- male and female mice. WT and NLRP3 -/- males and females challenged with PBS IP injection were used as negative control. All cells were CD45+, neutrophils were identified as Ly6C+/Ly6G+, macrophages were identified as Ly6C+/Ly6G-. Quantification of neutrophils in BAL from pristane-induced DAH WT and NLRP3 -/- male. WT and NLRP3 -/- male challenged with PBS IP injection were used as negative control. Results represent median +/- interquartile range, ANOVA multiple comparison test was used for statistical analysis between groups (ns: non-significant, p < 0.05^*, p < 0.01^**). (B) Lung tissue cryosections were obtained from pristane-induced DAH WT and NLRP3 -/- mice. WT and NLRP3 -/- mice challenged with PBS IP injection were a negative control: cryosections were immunostained for DNA (blue), Ly6G (white) and H3Cit (green). Bar 25µm. (C) Quantification of neutrophils in lungs from pristane-induced DAH WT and NLRP3 -/- mice. WT and NLRP3 -/- mice challenged with PBS IP injection were a negative control. Results represent median ± interquartile range, ANOVA multiple comparison test was used for statistical analysis between groups (ns: non-significant, p< 0.001^***). Quantification of H3Cit in lungs from pristane-induced DAH WT and NLRP3 -/- mice. WT and NLRP3 -/- mice challenged with PBS IP injection were a negative control. Results represent median ± interquartile range, ANOVA multiple comparison test was applied for statistical analysis between groups (ns: non-significant, p< 0.01^**, < 0.001^***). Each experiment had n=3-6 mice/PBS group and n=7-10 mice/pristane group.