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

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
ABSTRACT

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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Roche
Cocktail di inibitori di proteasi cOmplete, Mini, senza EDTA, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial
Sigma-Aldrich
Metile 4-idrossibenzoato, BioReagent, suitable for insect cell culture