Fig. 1
In vitro models of infection. Adhesion (a) and invasion (b) assays of individual clinical isolates from patients with acute pulmonary infections, bronchiectasis, or CF. Type II pneumocytes A549 cells monolayers were infected with 5 x105 CFU and the percentage of adhered and invaded cells was measured using a standard gentamycin exclusion assay. PAHM4 is highlighted on the graphs in red. Experiments were performed in triplicate. C. Macrophage uptake (solid bars, time = 15 min) and killing (striped bars, time = 90 min) were performed using murine bone marrow macrophages extracted from BALB/c mice and grown in presence of L-cells conditioned medium for 5 days. Macrophages were infected with PAO1, PA14 or PAHM4 at a MOI 500:1. Experiments were performed in duplicate. D. In vitro infection of lettuce leaves. The midrib of romaine lettuce was infected with a bacterial suspension of 106 CFU of PAO1, PA14 or PAHM4 and incubated at ambient temperature for 48 h. The diameter of the soft rot area at the site of inoculation was measured. Experiments were performed in triplicate. For all experiments, data were analyzed using an unpaired two-tailed t-test and the software GraphPad Prism 5.01