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. 2018 Aug 3;9:1733. doi: 10.3389/fmicb.2018.01733

FIGURE 5.

FIGURE 5

NADase prevents GAS-containing phagosome acidification in endothelial cells. (A) The intracellular localization of GAS (DAPI, blue), LC3 (Alexa 488, green), and acidification (acidotropic indicator LysoTracker Red DND-99, red) in endothelial cells were visualized by confocal microscopy after 1 and 5 h of infection. Representative images are shown from three independent experiments. Scale bar = 20 μm. (B) The percentage of intracellular GAS associated with LysoTracker was quantified at 1 and 5 h of infection. At least 100 intracellular GAS were quantified for each time point in at least three independent experiments. (C) The intracellular multiplication of NZ131 and nga mutant in endothelial cells after bafilomycin A1 treatment. HMEC-1 cells with or without pretreatment of bafilomycin A1 (100 nM) were infected with NZ131 or the nga mutant at M.O.I. of 1 and intracellular viable bacteria were counted by CFU-based assays. The data represent the means ± SEM of at least three independent experiments. ns, not significant; ∗∗p < 0.01; ∗∗∗p < 0.001 (two-way ANOVA).