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Quantification of Autophagy During Senescence

  • Joon Tae Park
  • Young-Sam Lee
  • Sang Chul Park
Part of the Methods in Molecular Biology book series (MIMB, volume 1896)

Abstract

Autophagy constitutes an evolutionarily conserved catabolic process that contributes to the clearance of damaged cellular components in response to a variety of stress conditions. Additionally, it plays a variety of physiological and pathophysiological roles in maintaining cell homeostasis. Recently, the critical role of autophagy during cellular senescence has been supported by evidences demonstrating the reversal of senescence by the reestablishment of autophagy. As considerable attention has been directed toward understanding the molecular mechanisms underlying senescence and autophagy, a method to accurately quantify autophagy during senescence is critical to understand its role in senescence and senescence-related diseases. In this chapter, we describe the use of CYTO-ID® green dye and DQ™ Red BSA to monitor the autophagic flux as an accurate method to quantify autophagic activity. This technique relies on the specificity of CYTO-ID® green dye in staining autophagosome and the cleavage of the self-quenched DQ™ Red BSA protease substrates in an acidic compartment. In particular, herein we describe protocols to quantify autophagy during senescence.

Key words

Autophagy Senescence CYTO-ID® green dye DQ™ Red BSA Autophagic flux 

Notes

Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2018R1D1A1B07040293), the DGIST R&D Program of the Ministry of Science, ICT and Technology of KOREA (2017010072 and 2017010115), and Chonnam National University R&D Program Grant for the Research Chair Professor.

Competing financial interests statement

The authors declare no competing financial interests.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Joon Tae Park
    • 1
  • Young-Sam Lee
    • 2
    • 3
  • Sang Chul Park
    • 2
    • 4
  1. 1.Division of Life Sciences, College of Life Sciences and BioengineeringIncheon National UniversityIncheonSouth Korea
  2. 2.Well Aging Research CenterDGISTDaeguSouth Korea
  3. 3.Department of New BiologyDGISTDaeguSouth Korea
  4. 4.The Future Life and Society Research CenterChonnam National UniversityGwangjuSouth Korea

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