Notch signaling assays in Drosophila cultured cell lines

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

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.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cell Line
  • Drosophila / cytology
  • Drosophila / genetics
  • Drosophila / metabolism*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Edetic Acid / metabolism
  • Egtazic Acid / metabolism
  • Gene Expression Regulation
  • Genes, Reporter
  • Luciferases / genetics
  • Polymerase Chain Reaction / methods
  • Receptors, Notch / genetics
  • Receptors, Notch / metabolism*
  • Renilla / enzymology
  • Signal Transduction*
  • Transfection

Substances

  • Drosophila Proteins
  • N protein, Drosophila
  • Receptors, Notch
  • Egtazic Acid
  • Edetic Acid
  • Luciferases