Abstract
The detection of autophagic vesicles in interphase cells is well characterized with markers such as LC3, SQSTM1 (also known as p62) and LAMP2, which are commonly used in immunofluorescence and biochemistry assays to evaluate the status of autophagy in adherent cells. During mitosis, cells undergo important morphological changes which alter the position of the central plane, therefore the imaging of dividing cells has to be specifically designed. Here, we describe a method to label and image autophagic vesicles in mitotic cells to systematically analyze their number, morphology and distribution.
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Abbreviations
- CDK1:
-
Cyclin-dependent kinase 1
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- DMEM:
-
Dulbecco’s modified eagle medium
- LAMP2:
-
Lysosome-associated membrane protein 2
- LC3:
-
Microtubule-associated proteins 1A/1B light chain 3
- PBS:
-
Phosphate-buffered saline
- PFA:
-
Paraformaldehyde
- ROI:
-
Region of interest
- SQSTM1:
-
Sequestosome-1
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Acknowledgments
This project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement no 799000 to C.M. We thank CERCA Program/Generalitat de Catalunya for institutional support.
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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Almacellas, E., Garcia-Cajide, M., Tauler, A., Mauvezin, C. (2022). Analysis of Autophagic Vesicles in Mitotic Cells. In: Norberg, H., Norberg, E. (eds) Autophagy and Cancer. Methods in Molecular Biology, vol 2445. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2071-7_9
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DOI: https://doi.org/10.1007/978-1-0716-2071-7_9
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-2071-7
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