Cells were seeded onto 35-mm-diameter tissue-coated petri dishes (35mm TC-Treated Culture Dish, Corning) with 2 ml of solution (347,000 cells/ml) and left in an incubator at 37°C and 5% CO2 for 24 hours. Cells were washed once with complete growth medium, and 2 ml of 10 nM QSH solutions was added. After 24 hours, petri dishes were removed from the incubator and washed twice with complete growth medium. LysoTracker Green (DND-26, Thermo Fisher Scientific) was added at 1 μM concentration, and confocal images were obtained. Lysosomal colocalization studies were performed using a spinning-disk confocal imaging system. Z-stacks were taken at 2-μm step sizes, with a total distance of 40 μm.

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