果蝇培养细胞系中的Notch信号通路检测

Notch signaling assays in Drosophila cultured cell lines.

作者信息

Li Jinghua, Housden Benjamin E, Bray Sarah J

机构信息

Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, CB2 3DY, Cambridge, UK.

出版信息

Methods Mol Biol. 2014;1187:131-41. doi: 10.1007/978-1-4939-1139-4_10.

DOI:10.1007/978-1-4939-1139-4_10

PMID:25053486

Abstract

Signaling assays in Drosophila cell lines are a valuable method for investigating whether other proteins influence the function of the Notch pathway and for assessing whether specific enhancers or genes are regulated by Notch. In this chapter, we will describe two different types of assays that can be used to monitor Notch activation in Kc167 and S2 cells. One involves activating Notch in cultured cells and measuring the change in endogenous gene expression levels. The other uses luciferase reporters and measures their response to Notch, by co-transfecting with NICD.

摘要

果蝇细胞系中的信号转导分析是一种有价值的方法，可用于研究其他蛋白质是否影响Notch信号通路的功能，以及评估特定增强子或基因是否受Notch调控。在本章中，我们将描述两种不同类型的分析方法，可用于监测Kc167和S2细胞中的Notch激活。一种方法是在培养细胞中激活Notch并测量内源性基因表达水平的变化。另一种方法是使用荧光素酶报告基因，并通过与NICD共转染来测量它们对Notch的反应。