Chapter title |
Assessing Autophagic Activity and Aggregate Formation of Mutant Huntingtin in Mammalian Cells
|
---|---|
Chapter number | 2 |
Book title |
Huntington’s Disease
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7825-0_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7824-3, 978-1-4939-7825-0
|
Authors |
Eleanna Stamatakou, Ye Zhu, David C. Rubinsztein, Stamatakou, Eleanna, Zhu, Ye, Rubinsztein, David C. |
Abstract |
The accumulation of mutant aggregate-prone proteins is a hallmark of the majority of neurodegenerative disorders, including Alzheimer's, Parkinson's, and Huntington's diseases. Autophagy, a cytosolic bulk degradation system, is the major clearance pathway for several aggregate-prone proteins, such as mutant huntingtin. The autophagosome-associated protein LC3-II is a specific marker of autophagic flux within cells, whereas aggregate formation of mutant huntingtin represents a good readout for studying autophagy modulation. Here we describe the method of assessing autophagic flux using LC3-II western blotting and substrate clearance by expressing the N-terminal fragment of huntingtin (htt exon 1) containing an expanded polyglutamine tract in mammalian cells. |
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