Sep 23, 2023
Mitophagy experiments
- Elias Adriaenssens1
- 1Sascha Martens lab, University of Vienna, Max Perutz Labs - Vienna
External link: https://doi.org/10.1038/s41594-024-01338-y
Protocol Citation: Elias Adriaenssens 2023. Mitophagy experiments. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj3yjnlk5/v1
Manuscript citation:
Adriaenssens, E., Nguyen, T.N., Sawa-Makarska, J. et al. Control of mitophagy initiation and progression by the TBK1 adaptors NAP1 and SINTBAD. Nat Struct Mol Biol 31, 1717–1731 (2024). https://doi.org/10.1038/s41594-024-01338-y
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 04, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 84439
Keywords: Mitophagy experiments, ASAPCRN, mitophagy in hela cell, mitophagy experiment, mitophagy experiments this protocol, mitophagy, hela cell, cell
Funders Acknowledgements:
Aligning Science Across Parkinson’s (ASAP)
Grant ID: ASAP-000350
Marie Skłodowska-Curie MSCA Postdoctoral fellowship
Grant ID: 101062916
Abstract
This protocol describes how to induce mitophagy in HeLa cells.
Attachments
772-1957.pdf
32KB
Materials
Reagents
Oligomycin AApexBio TechnologyCatalog #A5588
Antimycin A from Streptomyces sp.Merck MilliporeSigma (Sigma-Aldrich)Catalog #A8674
Q-VD-OPhApexBio TechnologyCatalog #A1901
Troubleshooting
Mitophagy experiments
8h
To induce mitophagy, treat cells with 10 micromolar (µM) Oligomycin and 4 micromolar (µM) Antimycin A.
In case cells are treated for more than 08:00:00 , add 10 micromolar (µM) Q-VD-OPh to suppress apoptosis.
8h
Analyze the samples by SDS-PAGE and western blot or flow cytometry.