|Cells were resuspended in 250 μL of phenol red-free culture medium (Thermo Fisher Scientific Inc.; No: 1294895) containing 5% FBS, and 250 μL of the diluted Cyto-ID® Green stain solution was added to each sample and mixed well. Cyto-ID® Green stain is a cationic amphiphilic tracer that selectively labels autophagic vacuoles in cells. Cells were incubated for 30 minutes at 37°C in the dark, collected by centrifugation, washed with 1× assay buffer, and resuspended in 500 μL fresh 1× assay buffer. Cells were subjected to flow cytometric analysis using the green (FL1) channel. The flow cytometer collected 10,000 events.
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