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Published on: August 27, 2019
Protocol for characterizing cell motility and extracellular signal-regulated kinase dynamics in epithelial monolayers
Julio Cesar Sanchez-Rendon1, Lara Hundsdorfer1, Effie E Bastounis1
1Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany; Cluster of Excellence EXC 2124 Controlling Microbes to Fight Infections, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany.
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Extracellular signal-regulated kinase (ERK) activity waves regulate critical processes like wound healing and bacterial pathogen dissemination by altering host cell motility and biomechanics. Here, we present a protocol for fluorescence resonance energy transfer (FRET) imaging of ERK activity in the nuclei of epithelial cells in a monolayer using an ERK biosensor. Moreover, we outline all image processing steps for the spatiotemporal quantification of single-cell ERK oscillations and wave propagation. Modifications, especially with respect to FRET imaging, may be necessary if different ERK biosensors are used. For complete details on the use and execution of this protocol, please refer to Hundsdorfer et al.1.
