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Detecting Autophagy and Autophagy Flux in Chronic Myeloid Leukemia Cells Using a Cyto-ID Fluorescence Spectrophotometric Assay

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Chronic Myeloid Leukemia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1465))

Abstract

Autophagy is a catabolic process whereby cellular components are degraded to fuel cells for longer survival during stress. Hence, autophagy plays a vital role in determining cell fate and is central for homeostasis and pathogenesis of many human diseases including chronic myeloid leukemia (CML). It has been well established that autophagy is important for the leukemogenesis as well as drug resistance in CML. Thus, autophagy is an intriguing therapeutic target. However, current approaches that detect autophagy lack reliability and often fail to provide quantitative measurements. To overcome this hurdle and facilitate the development of autophagy-related therapies, we have recently developed an autophagy assay termed as the Cyto-ID fluorescence spectrophotometric assay. This method uses a cationic fluorescence dye, Cyto-ID, which specifically labels autophagic compartments and is detected by a spectrophotometer to permit a large-scale and quantitative analysis. As such, it allows rapid, reliable, and quantitative detection of autophagy and estimation of autophagy flux. In this chapter, we further provide technical details of this method and step-by-step protocols for measuring autophagy or autophagy flux in CML cell lines as well as primary hematopoietic cells.

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Acknowledgment

We thank Yanping Liang and Susan Murphy in assisting us with our experiments. This work was supported by the start-up funds from the Virginia Tech Carilion Research Institute to Z.S.

Author information

Authors and Affiliations

  1. Virginia Tech Carilion Research Institute, Roanoke, VA, 24016, USA

    Sujuan Guo, Kevin J. Pridham & Zhi Sheng

  2. Graduate Program in Translational Biology, Medicine, and Health, Virginia Tech, Blacksburg, VA, 24061, USA

    Kevin J. Pridham

  3. Department of Biological Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, 24061, USA

    Zhi Sheng

  4. Department of Internal Medicine, Virginia Tech Carilion School of Medicine, Roanoke, VA, 24016, USA

    Zhi Sheng

  5. Faculty of Health Science, Virginia Tech, Blacksburg, VA, 24061, USA

    Zhi Sheng

Authors
  1. Sujuan Guo
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  2. Kevin J. Pridham
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  3. Zhi Sheng
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Corresponding author

Correspondence to Zhi Sheng .

Editor information

Editors and Affiliations

  1. Department of Medicine Division of Hematology/Oncology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Shaoguang Li

  2. Medical Research Institute, Wuhan University, Wuhan, China

    Haojian Zhang

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Guo, S., Pridham, K.J., Sheng, Z. (2016). Detecting Autophagy and Autophagy Flux in Chronic Myeloid Leukemia Cells Using a Cyto-ID Fluorescence Spectrophotometric Assay. In: Li, S., Zhang, H. (eds) Chronic Myeloid Leukemia. Methods in Molecular Biology, vol 1465. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4011-0_9

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  • DOI: https://doi.org/10.1007/978-1-4939-4011-0_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-4009-7

  • Online ISBN: 978-1-4939-4011-0

  • eBook Packages: Springer Protocols

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