Cells were seeded in 35-mm glass-bottomed dishes (MatTek Corporation) the day before the experiment in complete growth media (38). On the day of the experiment, cells were loaded with MitoTracker Red (MTR) (0.5 μM) for 60 min in a humidified atmosphere at 37°C in the growth media. Afterward, the cells were washed and incubated with LysoTracker Green (LTG) (0.5 μM) for 20 min. After MTR and LTG had been loaded, one-third of the initial concentration of LTG was kept in the media for the duration of the experiment. On the microscope stage, culture media (CM) (38) was replaced with CM plus 5 μM SoSe. Time series of live cell images were collected every 1 to 2 min for 150 min after addition of SoSe on the Olympus FluoView FV10i LIV laser scanning confocal microscope in the MUSC Cell and Molecular Imaging Core Facility.

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