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|Autophagosomes were detected with an autofluorescent compound monodansylcadaverine (MDC), using a Cell Meter Autophagy kit (AAT Bioquest, Inc.). Briefly, BMSCs were seeded on glass coverslips in a 24-wells plate at a density of 3 x 104/well. After incubation, BMSCs were stained with 1 x MDC solution and incubated at 37°C for 1 hour. Cells were washed three times and examined under a confocal microscope (Ex = 330 nm).|
|Incubate cells with CYTO-ID dye (2μl of CYTO-ID in 1ml of 1X assay buffer) for 30 min|
|Incase of Low CYTO-ID Green 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 Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experimen