Techniques for the Detection of Autophagy in Primary Mammalian Cells

Cold Spring Harbor Protocols
Daniel PulestonAnna Katharina Simon

Abstract

Autophagy is a lysosomal catabolic pathway responsible for the degradation of cytoplasmic constituents. Autophagy is primarily a survival pathway for recycling cellular material in times of nutrient starvation, and in response to hypoxia, endoplasmic reticulum stress, and other stresses, regulated through the mammalian target of rapamycin pathway. The proteasomal pathway is responsible for degradation of proteins, whereas autophagy can degrade cytoplasmic material in bulk, including whole organelles such as mitochondria (mitophagy), bacteria (xenophagy), or lipids (lipophagy). Although signs of autophagy can be present during cell death, it remains controversial whether autophagy can execute cell death in vivo. Here, we will introduce protocols for detecting autophagy in mammalian primary cells by using western blots, immunofluorescence, immunohistochemistry, flow cytometry, and imaging flow cytometry.

References

Oct 15, 2011·Autophagy·Daniel J KlionskySharon A Tooze
Nov 15, 2011·Cell·Noboru Mizushima, Masaaki Komatsu
Sep 12, 2012·Autophagy·Daniel J KlionskyBrian Zuckerbraun
Sep 4, 2014·Cold Spring Harbor Protocols·Lindsay DeVorkin, Sharon M Gorski
Aug 5, 2015·Cold Spring Harbor Protocols·Suganthi ChittaranjanSharon M Gorski
Sep 4, 2015·Cold Spring Harbor Protocols·Daniel Puleston

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Citations

Aug 9, 2016·Antiviral Research·Zoë R WallaceLucy Dorrell
Apr 21, 2017·Journal of the American Society of Nephrology : JASN·Tamadher A AlghamdiAndrew Advani
Jan 7, 2018·Biochemical Society Transactions·Kevin C YangSharon M Gorski
Dec 10, 2016·Cold Spring Harbor Protocols·John Silke, Ricky W Johnstone
Sep 4, 2015·Cold Spring Harbor Protocols·Kanchan Phadwal
Sep 4, 2015·Cold Spring Harbor Protocols·Daniel Puleston

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