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. 2013 Sep 18;8(9):e73111. doi: 10.1371/journal.pone.0073111

Figure 1. Colocalization of the P. aeruginosa exsA mutant with acidified vacuoles in epithelial cells compared to that of wild-type bacteria or a popB (translocon) mutant.

Figure 1

Confocal microscopy images of human corneal epithelial cells at 5 h post-infection with GFP-expressing P. aeruginosa (green). Prior to imaging, infected cultures were infused with LysoTracker (LT) DND-22 (pseudo-colored red). Panels depict (A) Uninfected control, (B) PAO1 exsA mutant (C) PAO1 popB (translocon) mutant and (D) wild-type PAO1. Uninfected cells appeared healthy. The intracellular exsA mutant appeared more frequently in LT (+) (acidified) vacuoles which co-localized yellow (arrows) than either the intracellular popB mutant or wild-type PAO1. PAO1-infected cells which displayed bleb-niche formation (1D inset) showed reduced fluorescence (< 10% fluorescence intensity of PAO1-infected non-blebbing cells, p < 0.001 Welch’s corrected t-Test). Occasional bleb-niches contained LT (+) vacuoles containing bacteria (1D inset, yellow). Representative images are shown. Magnification ~ 600 x.