Fig. 3.

Removal of Cif catalytic activity decreases recovered CFU counts and increases the amount of 15-epi-lipoxin A₄ (LXA₄) detected in an acute pneumonia model. A: immunoblot of Cif from PA14 and PA14-Cif_D129S in the absence (1:4 sample dilution) and presence (1:20 sample dilution) of an epoxide inducer. Blots generated at the same time and processed in parallel. B: ratio of recovered CFUs at 24 h postinfection to input CFUs from LH of mice treated with PA14 or PA14-Cif_D129S (n = 4 experiments, 4–6 mice for each treatment group/experiment; significance calculated using a 1-tailed exact Wilcoxon-Mann-Whitney test from the R package coin.). The gray box indicates those mice where no CFUs were detected. C: representative plots from BALF samples gated on live CD45⁺CD11b⁺ markers. Thin-lined black boxes indicate the CD11c⁺F4/80⁺ macrophage population, with the percent of parent gate indicated above the black box; thick-lined black boxes indicate the F4/80⁻CD11c⁻ population displayed in D. D: representative plots from BALF samples gated on live CD45⁺CD11b⁺CD11c⁻F4/80⁻ markers. Black inset boxes indicate the Ly6G⁺Ly6C^int neutrophil population, with the percent of parent gate indicated below the black box. E: counts of live CD45⁺CD11b⁺CD11c⁻F4/80⁻ macrophages/ml BALF. Not significant (ns), P > 0.05. F: counts of live CD45⁺CD11b⁺CD11c⁻F4/80⁻Ly6G⁺Ly6C^int neutrophils/ml BALF. G: percent of parent gates (live CD45⁺CD11b⁺CD11c⁻F4/80⁻) composed of neutrophils (Ly6G⁺Ly6C^int). E-G: n = 2 experiments, 4–6 mice for each treatment group/experiment; F and G: ns, P ≥ 0.2. H: background-subtracted levels of 15-epi LXA₄ detected in BALF from mice treated with PBS, PA14, or PA14-Cif_D129S (n = 4 experiments, 4–6 mice for each treatment group/experiment; ns, P ≥ 0.2.). E–H: black bars, medians; significance calculated using a Kruskal-Wallis Rank Sum test followed by a multiple-comparisons test in the R package “pgirmess.”