title: Co-culture Activation of MAP Kinase in Drosophila S2 Cells.
abstract: Intercellular communication often involves phosphorylation of signal transduction proteins, including mitogen-activated protein kinases (MAPKs). Immunological detection of phosphorylated MAPK can be used to monitor signaling in vivo, identify novel pathway components, and assess ligand activity. In this chapter, I describe a cell co-culture method to assess activity of cell-bound extracellular ligands that result in phosphorylation of the ERK (extracellular signal-regulated kinase) MAPK in Drosophila. This protocol may be adaptable to other pathways and/or model systems.
authors.affiliation: Department of Biology, Yeshiva University, 500 W. 185th St., New York, NY, 10033, USA. jsteinha@yu.edu.
authors.forename: Josefa
authors.initials: J
authors.lastname: Steinhauer
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D018920:Coculture Techniques; D004330:Drosophila / Q000378:metabolism*; D004789:Enzyme Activation; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D020928:Mitogen-Activated Protein Kinases
keywords: Co-culture; ERK; MAP kinase; S2 cells; Western blot; dp-ERK