Protocol for electrical modulation of ERK signaling dynamics in cell lines

Kan Zhu; Liang Guo; Preena Patel; John Albeck; Quan Qing; Wolfgang Losert; Min Zhao

STAR Protocols (Dec 2022)

Protocol for electrical modulation of ERK signaling dynamics in cell lines

Kan Zhu,
Liang Guo,
Preena Patel,
John Albeck,
Quan Qing,
Wolfgang Losert,
Min Zhao

Affiliations

Kan Zhu: Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA
Liang Guo: Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA; College of Intelligent Systems Science and Engineering, Harbin Engineering University, Harbin, Heilongjiang 150001, China
Preena Patel: Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA
John Albeck: Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA 95616, USA
Quan Qing: Department of Physics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA
Wolfgang Losert: Institute for Physical Science and Technology, Department of Physics, University of Maryland, College Park, MD 20742, USA
Min Zhao: Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 4
p. 101752

Abstract

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Summary: Here, we describe a protocol for modulating the dynamics of the extracellular signal-regulated kinases (ERK) pathway in a customized alternating current (AC) electric field stimulation chamber. We use an ERK translocation reporter that can accurately represent the intracellular ERK activity in real time without chemical agents or gene disruption. ERK activation is assessed by comparing the relative intensity of nuclear fluorescence to cytosolic fluorescence in live-cell conditions. The approach can be applied to other signaling pathways as well.For complete details on the use and execution of this protocol, please refer to Guo et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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