FIG. 5.

HAECs sense P. aeruginosa via TLR5. HAECs were seeded in 24-well plates. As described in Materials and Methods, 0.5 μg dominant negative human TLR5 (DNTLR5) (solid bars) or empty vector DNA (open bars) together with 0.3 μg of the hBD2 promoter-driven firefly luciferase gene and 10 ng of the Renilla luciferase reporter gene was used for each well. Cells were challenged 24 h after transfection with 10 μg/ml LTA or 100 CFU/cell of heat-killed PAO1 or PAK for 18 h, and then dual-luciferase assays were performed to measure induction of the luciferase activity. The data are means ± standard errors for at least two independent experiments. Samples were compared to a control. The group containing dominant negative human TLR5 (+DNTLR5) was compared to the group not containing dominant negative human TLR5 (−DNTLR5). An asterisk indicates that the P value is <0.001.