Methods for Monitoring ER-phagy

Marisa Di Monaco; Marián Péteri; Jin Rui Liang

doi:10.1007/978-1-0716-4067-8_9

Methods for Monitoring ER-phagy

Marisa Di Monaco
, Marián Péteri
, Jin Rui Liang (Lead / Corresponding author)

MRC PPU

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract

The endoplasmic reticulum (ER) serves as a central hub for protein synthesis, folding, and lipid biosynthesis in eukaryotic cells. Maintaining ER homeostasis is essential for optimal cellular function, and one mechanism that has garnered attention is endoplasmic reticulum-specific autophagy, or ER-phagy. ER-phagy selectively removes specific ER portions, playing a pivotal role in cellular health and adaptation to environmental stressors. ER-phagy can be induced by diverse cellular conditions such as amino acid starvation, disruption of ER quality control mechanisms, and accumulation of misfolded ER protein, highlighting cellular adaptability and the significance of ER-phagy in stress responses. Clinically relevant mutations in ER-phagy receptors are implicated in various diseases, underlining the fundamental importance of ER-phagy in ER homeostasis. Here, we provide comprehensive protocols and general considerations while investigating ER-phagy using three fundamental techniques—Western blotting, immunofluorescence, and flow cytometry—commonly used in ER-phagy detection and quantitation.

Original language	English
Title of host publication	Selective Autophagy
Subtitle of host publication	Methods and Protocols
Editors	Ioannis P. Nezis
Place of Publication	New York
Publisher	Humana Press
Chapter	9
Pages	109-126
Number of pages	18
Edition	1st
ISBN (Electronic)	9781071640678
ISBN (Print)	9781071640661, 9781071640692
DOIs	https://doi.org/10.1007/978-1-0716-4067-8_9
Publication status	Published - 2024

Publication series

Name	Methods in Molecular Biology
Volume	2845
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Autophagy
ER-phagy
Endoplasmic reticulum
FACS
Fluorescent reporters
Immunofluorescence
Selective autophagy
Western blotting

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-0716-4067-8_9

Cite this

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title = "Methods for Monitoring ER-phagy",

abstract = "The endoplasmic reticulum (ER) serves as a central hub for protein synthesis, folding, and lipid biosynthesis in eukaryotic cells. Maintaining ER homeostasis is essential for optimal cellular function, and one mechanism that has garnered attention is endoplasmic reticulum-specific autophagy, or ER-phagy. ER-phagy selectively removes specific ER portions, playing a pivotal role in cellular health and adaptation to environmental stressors. ER-phagy can be induced by diverse cellular conditions such as amino acid starvation, disruption of ER quality control mechanisms, and accumulation of misfolded ER protein, highlighting cellular adaptability and the significance of ER-phagy in stress responses. Clinically relevant mutations in ER-phagy receptors are implicated in various diseases, underlining the fundamental importance of ER-phagy in ER homeostasis. Here, we provide comprehensive protocols and general considerations while investigating ER-phagy using three fundamental techniques—Western blotting, immunofluorescence, and flow cytometry—commonly used in ER-phagy detection and quantitation. ",

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author = "\{Di Monaco\}, Marisa and Mari{\'a}n P{\'e}teri and Liang, \{Jin Rui\}",

note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.",

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PY - 2024

Y1 - 2024

N2 - The endoplasmic reticulum (ER) serves as a central hub for protein synthesis, folding, and lipid biosynthesis in eukaryotic cells. Maintaining ER homeostasis is essential for optimal cellular function, and one mechanism that has garnered attention is endoplasmic reticulum-specific autophagy, or ER-phagy. ER-phagy selectively removes specific ER portions, playing a pivotal role in cellular health and adaptation to environmental stressors. ER-phagy can be induced by diverse cellular conditions such as amino acid starvation, disruption of ER quality control mechanisms, and accumulation of misfolded ER protein, highlighting cellular adaptability and the significance of ER-phagy in stress responses. Clinically relevant mutations in ER-phagy receptors are implicated in various diseases, underlining the fundamental importance of ER-phagy in ER homeostasis. Here, we provide comprehensive protocols and general considerations while investigating ER-phagy using three fundamental techniques—Western blotting, immunofluorescence, and flow cytometry—commonly used in ER-phagy detection and quantitation.

AB - The endoplasmic reticulum (ER) serves as a central hub for protein synthesis, folding, and lipid biosynthesis in eukaryotic cells. Maintaining ER homeostasis is essential for optimal cellular function, and one mechanism that has garnered attention is endoplasmic reticulum-specific autophagy, or ER-phagy. ER-phagy selectively removes specific ER portions, playing a pivotal role in cellular health and adaptation to environmental stressors. ER-phagy can be induced by diverse cellular conditions such as amino acid starvation, disruption of ER quality control mechanisms, and accumulation of misfolded ER protein, highlighting cellular adaptability and the significance of ER-phagy in stress responses. Clinically relevant mutations in ER-phagy receptors are implicated in various diseases, underlining the fundamental importance of ER-phagy in ER homeostasis. Here, we provide comprehensive protocols and general considerations while investigating ER-phagy using three fundamental techniques—Western blotting, immunofluorescence, and flow cytometry—commonly used in ER-phagy detection and quantitation.

KW - Autophagy

KW - ER-phagy

KW - Endoplasmic reticulum

KW - FACS

KW - Fluorescent reporters

KW - Immunofluorescence

KW - Selective autophagy

KW - Western blotting

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M3 - Chapter

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SN - 9781071640661

SN - 9781071640692

T3 - Methods in Molecular Biology

SP - 109

EP - 126

BT - Selective Autophagy

A2 - Nezis, Ioannis P.

PB - Humana Press

CY - New York

ER -

Methods for Monitoring ER-phagy

Abstract

Publication series

Keywords

ASJC Scopus subject areas

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A CRISPR view on autophagy

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