Abstract
Single-cell analysis of the mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase (ERK) provides a means to perform highly detailed kinetic studies, assess heterogeneity between cells, and distinguish the subcellular localization of ERK activity. We describe here the methods needed to perform such measurements in a cell type of the investigator’s choosing. We discuss the selection of appropriate reporters and provide detailed methods for stably introducing reporters, collecting live-cell data, and automatically extracting quantitative information from individual cells.
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Acknowledgments
We thank Didem Sarikaya for helpful feedback on the manuscript. The methods described here were developed in part through support from the American Cancer Society (IRG-95-125-16).
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Pargett, M., Gillies, T.E., Teragawa, C.K., Sparta, B., Albeck, J.G. (2017). Single-Cell Imaging of ERK Signaling Using Fluorescent Biosensors. In: Tan, AC., Huang, P. (eds) Kinase Signaling Networks. Methods in Molecular Biology, vol 1636. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7154-1_3
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DOI: https://doi.org/10.1007/978-1-4939-7154-1_3
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