FIG. 5.

Minimal activity of ExoU-LINKER molecules affect the actin cytoskeleton of infected cells. A549 cell monolayers were infected with PA103ΔUT strains at an MOI of 10 for 4 h. After infection, samples were permeabilized and Texas Red-X phalloidin was used to visualize the actin cytoskeleton. Polyclonal anti-LPS antibody, followed by a secondary goat anti-rabbit Alexa488-labeled antibody, was used to detect P. aeruginosa. Slides were examined at ×20 and ×60 magnification of an objective lens to observe phalloidin staining of the actin cytoskeleton (left, ×20), in conjunction with the presence of the bacteria (right, ×60). The scale bars for ×20 and ×60 magnification are 50 and 10 μm, respectively. A549 cells infected with PA103ΔUT ExoT served as a positive control for translocation and cell rounding.