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. 2024:2845:109-126.
doi: 10.1007/978-1-0716-4067-8_9.

Methods for Monitoring ER-Phagy

Marisa Di Monaco  1 , Marian Peteri  1 , Jin Rui Liang  2
Affiliations

Methods for Monitoring ER-Phagy

Marisa Di Monaco et al. Methods Mol Biol. 2024.

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.

Keywords: Autophagy; ER-phagy; Endoplasmic reticulum; FACS; Fluorescent reporters; Immunofluorescence; Selective autophagy; Western blotting.

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References

    1. Khaminets A, Heinrich T, Mari M, Grumati P, Huebner AK, Akutsu M, Liebmann L, Stolz A, Nietzsche S, Koch N et al (2015) Regulation of endoplasmic reticulum turnover by selective autophagy. Nature 522:354–358. https://doi.org/10.1038/nature14498 - DOI - PubMed
    1. Grumati P, Morozzi G, Hölper S, Mari M, Harwardt M-LI, Yan R, Müller S, Reggiori F, Heilemann M, Dikic I (2017) Full length RTN3 regulates turnover of tubular endoplasmic reticulum via selective autophagy. eLife 6:e25555. https://doi.org/10.7554/eLife.25555 - DOI - PubMed - PMC
    1. Chino H, Hatta T, Natsume T, Mizushima N (2019) Intrinsically disordered protein TEX264 mediates ER-phagy. Mol Cell 74:909–921.e6. https://doi.org/10.1016/j.molcel.2019.03.033 - DOI - PubMed
    1. An H, Ordureau A, Paulo JA, Shoemaker CJ, Denic V, Harper JW (2019) TEX264 is an endoplasmic reticulum-resident ATG8-interacting protein critical for ER remodeling during nutrient stress. Mol Cell 74:891–908.e10. https://doi.org/10.1016/j.molcel.2019.03.034 - DOI - PubMed - PMC
    1. Smith MD, Harley ME, Kemp AJ, Wills J, Lee M, Arends M, von Kriegsheim A, Behrends C, Wilkinson S (2018) CCPG1 Is a non-canonical autophagy cargo receptor essential for ER-phagy and pancreatic ER proteostasis. Dev Cell 44:217–232.e11. https://doi.org/10.1016/j.devcel.2017.11.024 - DOI - PubMed - PMC

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