Fig. 5

Appearance of leukocytes in CNS during PA-induced lung infection. a White blood counts in control and PA-infected animals (OD₆₀₀ 0.3). Mann–Whitney test, P = 0.016 vs. control (uninfected) group. b Neutrophil and monocyte counts in blood obtained from control and PA-infected mice (OD₆₀₀ 0.3) measured by ProCyte One hematology analyzer. Mann–Whitney test, P = 0.04, P = 0.03 vs. control (uninfected) group. c Flow cytometry analysis of brain tissue homogenates labeled with anti-CD45 and anti-CD11b antibodies with gating on CD45⁺ cells. Quantitative analyses of (d) infiltrated leukocytes and (e) microglia in control and PA infected mice. Mann–Whitney test, P = 0.03 vs. control (uninfected) group. In all graphs, each point indicates data from an individual animal, bar graphs (columns and error bars) show mean ± S.E.M. f Representative confocal images of infiltrated leukocytes (CD45⁺; green) in the brain from a control (left) and a PA-lung infected mouse (right). DAPI (nuclei; blue). g GFAP immunoblotting in brain homogenates from control and PA-infected (24 h after infection) mice, and quantification of GFAP protein abundance relative to β-actin loading control. Mann–Whitney test, P = 0.016 vs. control (uninfected) group. h Representative confocal images of GFAP immunodetection (red), lectin-positive brain endothelial cells (green) and DAPI (nuclei; blue) in the brain of a control (left) and a PA-lung infected mouse (right). The inset in each main image (control and infected) represents a closer view of the indicated region at a higher magnification (right panels). In all graphs, each point indicates data from an individual animal, bar graphs (columns and error bars) show mean ± S.E.M