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Co-culture Activation of MAP Kinase in Drosophila S2 Cells

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ERK Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1487))

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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.

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Acknowledgements

I thank Jean-Yves Roignant and Kevin Legent for help developing the ERK activation assay; Benny Shilo for the D2F cells; Jessica Treisman for support during the development of the protocol and critical comments on the manuscript. This work was supported by the March of Dimes Birth Defects Foundation (to Jessica Treisman) and the National Institutes of Health (grant number EY13777 to Jessica Treisman from the National Eye Institute).

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Authors and Affiliations

  1. Department of Biology, Yeshiva University, 500 W. 185th St., New York, NY, 10033, USA

    Josefa Steinhauer

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  1. Josefa Steinhauer
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Correspondence to Josefa Steinhauer .

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Editors and Affiliations

  1. Institut de Biologia Molecular de Barcelona-CSIC, Parc Científic de Barcelona, Barcelona, Spain

    Gerardo Jimenez

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Steinhauer, J. (2017). Co-culture Activation of MAP Kinase in Drosophila S2 Cells. In: Jimenez, G. (eds) ERK Signaling. Methods in Molecular Biology, vol 1487. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6424-6_17

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  • DOI: https://doi.org/10.1007/978-1-4939-6424-6_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6422-2

  • Online ISBN: 978-1-4939-6424-6

  • eBook Packages: Springer Protocols

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