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|Five GC cell lines (AGS, N87, NCC19, NCC59 and SNU1967) were grown under the conditions described above. For assaying autophagy, a Cyto-ID Autophagy detection kit (Enzo Life Sciences, Farmingdale, New York, USA) was used according to the manufacturer's protocol. For assaying apoptosis, an Annexin V: PE Apoptosis detection Kit I (BD Biosciences, Franklin Lakes, New Jersey, USA) was used according to the manufacturer's protocol. FACSVerse (BD Biosciences) was used for flow cytometry analysis.|
|Use Cyto-ID dye (1 μl/4 ml assay buffer) for 30 minutes in dark at RT.|
|Incase of Low CYTO-ID dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
Incase of High CYTO-ID staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment