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

FIGURE 4.

FIGURE 4

NADase prevents GAS-containing vacuole trafficking to lysosomes in endothelial cells. (A) The intracellular localization of GAS, LC3, and lysosome in endothelial cells after 1 and 5 h of infection. Confocal microscopy was used to observe the co-localization of GAS (DAPI, blue) with LC3 (Alexa 488, green) and LAMP-1 (Alexa 594, red) in HMEC-1 cells. Representative images are shown from three independent experiments. Scale bar = 20 μm. (B) The intracellular GAS associated with LAMP-1 at 1 and 5 h of infection were quantified from confocal microscopy images. At least 100 intracellular GAS were quantified for each time point in at least three independent experiments. The data represent the means ± SEM of at least three independent experiments. ns, not significant; ∗p < 0.05 (two-way ANOVA). (C) The LC3 conversion in endothelial cells after infection with NZ131 or the nga mutant. HMEC-1 cell lysates from uninfected (cell alone) or infected cells and the LC3 conversion were determined by the western blotting. (D) The expression of LC3-I and LC3-II were measured by densitometry analysis and was expressed as the ratio of LC3-II/LC3-I.