Jan 12, 2026
Mitophagy assay
- Yongjia Duan1
- 1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Protocol Citation: Yongjia Duan 2026. Mitophagy assay. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp13r8gzp/v1
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: January 12, 2026
Last Modified: January 12, 2026
Protocol Integer ID: 238513
Keywords: ASAPCRN, mitophagy, assay
Funders Acknowledgements:
Michael J. Fox Foundation
Grant ID: ASAP-000350
Abstract
Mitophagy assay
Troubleshooting
Mitophagy assay
About 0.5 x10^5 cells stably expressing mito‐QC mitophagy reporter system (mCherry‐GFP‐FIS1101–152)
were seeded onto sterile glass coverslips in 24‐well dishes.
Next day, treated cell with or without 0.5mM DFP for 24 hours
After treatment, coverslips were washed twice with PBS, fixed with 3.7% (w/v)
formaldehyde, 200 mM HEPES pH 7.0 for 10 min and washed twice with PBS
After nuclei counterstaining with 1 μg/ml Hoechst‐33258 dye, slides were washed and
mounted in ProLong Gold (Invitrogen).
Image were captured with Nikon A1 confocal microscope under a 60 x oil objective.