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The Detection Techniques for Autophagy-Associated Cell Death-Related Genes and Proteins: Gene Expression Assay and Immunohistochemistry

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Autophagy in Differentiation and Tissue Maintenance

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1854))

Abstract

Autophagy is important in cellular homeostasis for the cell survival mechanism. Deficiency or excess of autophagy is generally related to some of diseases such as cancer and neurodegeneration. Although autophagy is a cell survival mechanism, it can mediate programmed cell death in several conditions. Autophagy-related genes (ATGs) regulate the autophagy and also control the crosstalk with autophagy-associated cell death and apoptosis in some condition. Various methods have been used to detect the marker genes and the proteins involved in these processes. Quantitative real-time PCR (qRT-PCR) method for monitoring the expression of genes involved in autophagy or autophagic cell death is often preferred because of its sensitivity, high efficiency potential, accurate quantification, and high-grade potential automation. The detection of the markers for autophagy-related process by immunohistochemistry in paraffin sections of various patient tissues has become a reliable method for monitoring autophagy. Here, we introduce protocols for detecting autophagy and autophagy-associated cell death in HeLa cells by using gene expression assays qRT-PCR, and also in paraffin-embedded tissue section from human biopsy material by using immunohistochemistry.

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Acknowledgment

This study was supported by the Research Fund of the University of Istanbul, Turkey, Project numbers: 17492, 38781 and 39211.

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Authors and Affiliations

  1. Medical Biology Department, Cerrahpasa Faculty of Medicine, Istanbul University, Cerrahpasa, Istanbul, 34098, Turkey

    Melek Ozturk & Fatma Kaya Dagistanli

  2. Department of Radiobiology, Institute of Graduate Studies in Science and Engineering, Istanbul University, Istanbul, Turkey

    Ozlem Dagdeviren Ozsoylemez

Authors
  1. Melek Ozturk
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  2. Ozlem Dagdeviren Ozsoylemez
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  3. Fatma Kaya Dagistanli
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Corresponding author

Correspondence to Melek Ozturk .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, ON, Canada

    Kursad Turksen

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© 2017 Springer Science+Business Media New York

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Ozturk, M., Ozsoylemez, O.D., Dagistanli, F.K. (2017). The Detection Techniques for Autophagy-Associated Cell Death-Related Genes and Proteins: Gene Expression Assay and Immunohistochemistry. In: Turksen, K. (eds) Autophagy in Differentiation and Tissue Maintenance. Methods in Molecular Biology, vol 1854. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_67

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  • DOI: https://doi.org/10.1007/7651_2017_67

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8747-4

  • Online ISBN: 978-1-4939-8748-1

  • eBook Packages: Springer Protocols

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