Fig. 2.

VopQ disrupts acidification in yeast vacuoles and HeLa lysosomes. (A) Proton translocation activity of vacuoles was measured as described (SI Materials and Methods) in the presence of 100 nM bafilomycin or increasing concentrations of His₆-VopQ. (B) HeLa cells expressing GFP-LC3 were microinjected with 10 µM GST or His₆-VopQ, incubated 10 min (37 °C, 5% CO₂), and then incubated with 75 nM LysoTracker Red for 10 min and visualized. (Scale bar, 20 µm.) (C) HeLa cells expressing GFP-LC3 were incubated with or without 25 μM chloroquine for 30 min and then either treated with 100 μM rapamycin or infected with the noted V. parahaemolyticus strains at a multiplicity of infection of 10 for 3 h. Cell lysates were separated via SDS/PAGE and immunoblotted for GFP and tubulin.