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Protocol for evaluating autophagy using LysoTracker staining in the epithelial follicle stem cells of the Drosophila ovary.

STAR protocols (2021-06-26)
Iliana Correa, Melissa Wang, Eric H Lee, Dara M Ruiz-Whalen, Alana M O'Reilly, Tanu Singh
RESUMEN

We have outlined the approach of visualizing autophagy specifically in the epithelial follicle stem cells of the Drosophila ovary using the LysoTracker dye. The advantage of using this protocol is that it details several techniques, including ovary dissection, immunofluorescence, and western blotting, that positively identify autophagy changes in a very small population of cells. One of the limitations of this protocol is that it needs to be combined with other genetic manipulations and positive markers of the autophagy pathway. For complete details on the use and execution of this protocol, please refer to Singh et al., (2018).

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Metil 4-hidroxibenzoato, BioReagent, suitable for insect cell culture