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

Abstract

Summary 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) .

Bioinformatics-based Investigation of Autophagy-Related Biomarkers in Heritable Ovarian Carcinoma

“…Studies have reported the potential of IL24 as a diagnostic marker in ovarian tumor subtypes, supporting its relevance in subtype-speci c diagnosis [17,18]. PTK6, also known as Protein Tyrosine Kinase 6, is another gene of interest that has been implicated in cancer pathogenesis [19][20][21]. Previous investigations have demonstrated the correlation between PTK6 expression and cancer stages in ovarian carcinoma, suggesting its potential as a staging biomarker for disease prognosis and management [22,23].…”

Sequential events during the quiescence to proliferation transition establish patterns of follicle cell differentiation in the Drosophila ovary

“…All fly stocks were raised on standard fly food (7.5 g/l agar, 83.6 g/l cornmeal, 50 ml/l molasses, 20 g/l yeast, 5.2 ml/l propionic acid, 10 ml tegosept/l). Nutrient restriction was accomplished by placing flies in collection cages on grape juice plates (50% grape juice, 1% acetic acid, 3% Bacto-Agar, 0.1% methylparaben in water; Correa et al, 2021 ) for a minimum of 3 days ( Hartman et al, 2015 ). Note that molasses plates do not induce quiescence in FSCs ( Ables et al, 2012 ; Hartman et al, 2013 , 2015 ) and are thus not appropriate for nutrient restriction conditions needed to analyze Q→P transitions.…”

The ERK1/2-Elk1, JNK-cJun, and JAK-STAT Transcriptional Axes as Potential Bortezomib Resistance Mediators in Prostate Cancer

“…To monitor the autophagic activity, the lysosomal marker Lysotracker RED was used. Lysotracker has been used as an autophagy marker, even though it does not directly assess autophagic flux but rather stains and thus quantifies acidic proteins (58)(59)(60). The acidic protein content of a cell is correlated to the lysosomal load, and therefore, due to the organelles' role in autophagy, Lysotracker can be used to study autophagic activity.…”