Abstract
The autophagosome-associated protein LC3-II is commonly used as a marker of autophagic activity within cells, but its levels are affected by both formation and degradation of autophagosomes. This can make the significance of altered LC3-II levels ambiguous. Here we describe the method of Bafilomycin A1 blotting, in which the degradation of autophagosomes is prevented in cultured cells, allowing the causes of altered LC3-II levels to be determined.
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Acknowledgements
We are grateful for funding from the Wellcome Trust to D.C.R. (Principal Research Fellowship) and A.S. (PhD Studentship).
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Streeter, A., Menzies, F.M., Rubinsztein, D.C. (2016). LC3-II Tagging and Western Blotting for Monitoring Autophagic Activity in Mammalian Cells. In: Castrillo, J., Oliver, S. (eds) Systems Biology of Alzheimer's Disease. Methods in Molecular Biology, vol 1303. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2627-5_8
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DOI: https://doi.org/10.1007/978-1-4939-2627-5_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2626-8
Online ISBN: 978-1-4939-2627-5
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