P aeruginosa induces neutrophil extravasation and migration. Neutrophils migrate significantly more to a source of P aeruginosa compared with iEC media alone. (A) Representative images of neutrophils migrating at 4-hour intervals. Neutrophils stained with calcein AM. Gradient direction shown in red. White line indicates the edge of the endothelial lumen. Scale bar, 100 µm. (B) The number of neutrophils outside the lumen was counted at 1-hour intervals, using particle analysis in FIJI. (C) The distance from the lumen edge was measured for all in-focus neutrophils outside the lumen at 4-hour intervals. Each bar represents the mean plus standard error of the mean (SEM). (B-C) Data quantified from 9 lumens per condition across 3 independent experiments. Asterisks represents significance between conditions at each point. ****P < .0001.