title: Identifying direct Notch transcriptional targets using the GSI-washout assay.
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.
authors: Department of Pathology and Laboratory Medicine, Abramson Family Cancer Research Institute, and Institute for Immunology, The Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 19104, USA.
authors: Will
authors: W
authors: Bailis
authors: Yumi
authors: Y
authors: Yashiro-Ohtani
authors: Warren S
authors: WS
authors: Pear
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D006801:Humans; D051880:Receptors, Notch / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000187:drug effects; D015533:Transcriptional Activation
chemical_list: D051880:Receptors, Notch; D053829:Amyloid Precursor Protein Secretases