Chapter title |
Measuring autophagy in stressed cells.
|
---|---|
Chapter number | 10 |
Book title |
Stress Responses
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2522-3_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2521-6, 978-1-4939-2522-3
|
Authors |
Marina N Sharifi, Erin E Mowers, Lauren E Drake, Kay F Macleod, Sharifi MN, Mowers EE, Drake LE, Macleod KF, Marina N. Sharifi, Erin E. Mowers, Lauren E. Drake, Kay F. Macleod |
Abstract |
Macro-autophagy is a major catabolic process in the cell used to degrade protein aggregates, dysfunctional organelles and intracellular pathogens that would otherwise become toxic. Autophagy also generates energy and metabolites for the cell through recycling of degraded autophagosomal cargo, which can be particularly important for cell viability under stress. The significance of changes in the rates of autophagic flux for cellular function and disease is being increasingly appreciated, and interest in measuring autophagy in different experimental systems is growing accordingly. Here, we describe key methodologies used in the field to measure autophagic flux, including monitoring LC3 processing by western blot, fluorescent cell staining, and flow cytometry, in addition to changes in the levels or posttranslational modifications of other autophagy markers, such as p62/Sqstm1 and the Atg5-Atg12 conjugate. We also describe what cellular stresses may be used to induce autophagy and how to control for changes in the rates of autophagic flux as opposed to inhibition of flux. Finally, we detail available techniques to monitor autophagy in vivo. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United States | 3 | 60% |
Unknown | 2 | 40% |
Demographic breakdown
Type | Count | As % |
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Scientists | 2 | 40% |
Members of the public | 2 | 40% |
Practitioners (doctors, other healthcare professionals) | 1 | 20% |
Mendeley readers
Geographical breakdown
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Unknown | 101 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 23 | 23% |
Researcher | 18 | 18% |
Student > Bachelor | 16 | 16% |
Student > Master | 10 | 10% |
Student > Doctoral Student | 5 | 5% |
Other | 8 | 8% |
Unknown | 21 | 21% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 26 | 26% |
Agricultural and Biological Sciences | 14 | 14% |
Medicine and Dentistry | 12 | 12% |
Pharmacology, Toxicology and Pharmaceutical Science | 8 | 8% |
Neuroscience | 6 | 6% |
Other | 12 | 12% |
Unknown | 23 | 23% |