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Lookup NU author(s): Dr Tetsushi Kataura, Professor Viktor KorolchukORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2023 The Author(s)Autophagy, a catabolic process governing cellular and energy homeostasis, is essential for cell survival and human health. Here, we present a protocol for generating autophagy-deficient (ATG5−/−) human neurons from human embryonic stem cell (hESC)-derived neural precursors. We describe steps for analyzing loss of autophagy by immunoblotting. We then detail analysis of cell death by luminescence-based cytotoxicity assay and fluorescence-based TUNEL staining. This hESC-based experimental platform provides a genetic knockout model for undertaking autophagy studies relevant to human biology. For complete details on the use and execution of this protocol, please refer to Sun et al. (2023).1
Author(s): Korsgen ME, Sun C, Seranova E, Zatyka M, Astuti D, Kataura T, Barrett T, Korolchuk VI, Sarkar S
Publication type: Article
Publication status: Published
Journal: STAR Protocols
Year: 2023
Volume: 4
Issue: 3
Print publication date: 15/09/2023
Online publication date: 25/08/2023
Acceptance date: 02/04/2018
Date deposited: 13/11/2023
ISSN (electronic): 2666-1667
Publisher: Cell Press
URL: https://doi.org/10.1016/j.xpro.2023.102529
DOI: 10.1016/j.xpro.2023.102529
Data Access Statement: This protocol does not involve or generate any specific dataset or code.
PubMed id: 37624702
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