Cell Cycle Analysis of ER Stress and Autophagy

A Popat; A A Patel; Gary Warnes

doi:10.1007/978-1-0716-2553-8_13

Cell Cycle Analysis of ER Stress and Autophagy

Methods Mol Biol. 2022:2543:155-166. doi: 10.1007/978-1-0716-2553-8_13.

Authors

A Popat¹, A A Patel¹, Gary Warnes²

Affiliations

¹ Flow Cytometry Core Facility, The Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary London University, London, UK.
² Flow Cytometry Core Facility, The Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary London University, London, UK. g.warnes@qmul.ac.uk.

PMID: 36087266
DOI: 10.1007/978-1-0716-2553-8_13

Abstract

Autophagy and ER stress are most often studied employing a Western blotting approach to the measurement of autophagy by LC3B upregulation and the ER stress sensor signaling proteins PERK (protein kinase R-like endoplasmic reticulum kinase), IRE1, and ATF6 which initiate protein refolding and elongation of the ER until ER homeostasis is returned. If the misfolding of proteins is increased, then ER stress is maintained, and microautophagy of the ER or specifically reticulophagy occurs. However, LC3B, PERK, protein misfolding, and changes in ER mass (reticulophagy) can also be measured in a cell cycle-dependent manner by flow cytometry and the use of antibodies, protein misfolding, and ER tracking fluorescent probes.

Keywords: Autophagy; ER stress; Misfolded proteins; PERK; Reticulophagy.

MeSH terms

Autophagy
Cell Cycle
Endoplasmic Reticulum / metabolism
Endoplasmic Reticulum Stress*
eIF-2 Kinase* / metabolism

Substances

eIF-2 Kinase