Assessment of EGFP-Q74 degradation for the measurement of autophagic flux

Yan Wang; Jiao Meng; Qi Wu; Guo Chen; Wei Xie; Zhen Zhang; Guido Kroemer; Oliver Kepp

doi:10.1016/bs.mcb.2020.10.018

Assessment of EGFP-Q74 degradation for the measurement of autophagic flux

Yan Wang, Jiao Meng, Qi Wu, Guo Chen, Wei Xie, Zhen Zhang, Guido Kroemer, Oliver Kepp

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

1 Citation (Scopus)

Abstract

Autophagy plays a major role in physiological and pathological processes. The quantitation of the abundance of autophagy-specific substrates constitutes an efficient strategy for assessing autophagic activity. Here, we provide a detailed protocol for quantifying the decay of a fusion protein composed by enhanced green fluorescent protein (EGFP) and glutamine repeats (Q74) using regular or high-throughput fluorescence microscopy. This method provides a direct measurement of autophagic flux in a Huntington's disease model.

Original language	English
Title of host publication	Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B
Editors	Oliver Kepp, Lorenzo Galluzzi
Publisher	Academic Press Inc.
Pages	31-38
Number of pages	8
ISBN (Print)	9780128244876
DOIs	https://doi.org/10.1016/bs.mcb.2020.10.018
Publication status	Published - 1 Jan 2021

Publication series

Name	Methods in Cell Biology
Volume	165
ISSN (Print)	0091-679X

Keywords

Autophagy
EGFP-Q74
Fluorescence microscopy
Huntington's disease

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10.1016/bs.mcb.2020.10.018

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Wang, Y., Meng, J., Wu, Q., Chen, G., Xie, W., Zhang, Z., Kroemer, G., & Kepp, O. (2021). Assessment of EGFP-Q74 degradation for the measurement of autophagic flux. In O. Kepp, & L. Galluzzi (Eds.), Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B (pp. 31-38). (Methods in Cell Biology; Vol. 165). Academic Press Inc.. https://doi.org/10.1016/bs.mcb.2020.10.018

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Assessment of EGFP-Q74 degradation for the measurement of autophagic flux. / Wang, Yan; Meng, Jiao; Wu, Qi et al.
Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B. ed. / Oliver Kepp; Lorenzo Galluzzi. Academic Press Inc., 2021. p. 31-38 (Methods in Cell Biology; Vol. 165).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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T1 - Assessment of EGFP-Q74 degradation for the measurement of autophagic flux

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AU - Zhang, Zhen

AU - Kroemer, Guido

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AB - Autophagy plays a major role in physiological and pathological processes. The quantitation of the abundance of autophagy-specific substrates constitutes an efficient strategy for assessing autophagic activity. Here, we provide a detailed protocol for quantifying the decay of a fusion protein composed by enhanced green fluorescent protein (EGFP) and glutamine repeats (Q74) using regular or high-throughput fluorescence microscopy. This method provides a direct measurement of autophagic flux in a Huntington's disease model.

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PB - Academic Press Inc.

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Assessment of EGFP-Q74 degradation for the measurement of autophagic flux

Abstract

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Keywords

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