Chapter title |
Identifying Direct Notch Transcriptional Targets Using the GSI-Washout Assay
|
---|---|
Chapter number | 19 |
Book title |
Notch Signaling
|
Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-1139-4_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1138-7, 978-1-4939-1139-4
|
Authors |
Will Bailis, Yumi Yashiro-Ohtani, Warren S. Pear, Bailis, Will, Yashiro-Ohtani, Yumi, Pear, Warren S. |
Abstract |
Genetic gain- and loss-of-function studies have traditionally been used to study transcriptional networks regulated by the Notch signaling pathway; however these techniques lack the ability to resolve primary and secondary transcriptional events. In contrast, the γ-secretase inhibitor (GSI) washout assay takes advantage of the reversibility of GSI, a pharmacological inhibitor of Notch signaling, along with the ability of cycloheximide to prevent secondary transcriptional effects to identify direct Notch pathway targets. Here we review this technique and the technical considerations for adapting this assay to a cell type of choice. |
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