FIGURE 6. atg7^−/− mice exhibited exacerbated lung infection and bacterial burdens against Pa.

WT mice and atg7^−/− mice were infected with 1×10⁷ CFU of PAO1. After 24 h, AMs were isolated from BAL fluids. (A) Cell viability was tested by MTT assay. (B) Mitochondrial potential was assessed by the JC-1 fluorescence assay. (C) Superoxide production was determined using H₂DCF assay. (D) H₂O₂ production was determined using EuTc assay. (E) NO production was detected using Griess reagent. (F) The lung homogenates were used for immunoblotting to determine the relevant signaling pathways and phosphorylation of signaling proteins. (G) Lungs were removed for histological analysis (inset showing the typical tissue injury and inflammatory influx). (H) The leukocyte infiltration score in lungs was determined by blindly (for the samples) enumerating the lymphocytes. (I) Lungs were homogenized in PBS and were used for assessing bacterial colonies. Data are shown as means±SEM from three independent experiments. (One-way ANOVA (Tukey’s post hoc); *, p<0.05; **, p<0.01).