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

doi:10.1016/j.xpro.2022.101752

Protocol for electrical modulation of ERK signaling dynamics in cell lines

STAR Protoc. 2022 Dec 16;3(4):101752. doi: 10.1016/j.xpro.2022.101752. Epub 2022 Oct 7.

Authors

Kan Zhu¹, Liang Guo², Preena Patel¹, John Albeck³, Quan Qing⁴, Wolfgang Losert⁵, Min Zhao⁶

Affiliations

¹ Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA.
² 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.
³ Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA 95616, USA.
⁴ Department of Physics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
⁵ Institute for Physical Science and Technology, Department of Physics, University of Maryland, College Park, MD 20742, USA.
⁶ Department of Ophthalmology & Vision Science, Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, Sacramento, CA 95817, USA. Electronic address: minzhao@ucdavis.edu.

Abstract

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).

Keywords: Biotechnology and bioengineering; Cell biology; Cell culture; Microscopy; Signal transduction.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Line
Extracellular Signal-Regulated MAP Kinases* / metabolism
MAP Kinase Signaling System
Phosphorylation
Signal Transduction*

Substances

Extracellular Signal-Regulated MAP Kinases

Grants and funding

R01 EY019101/EY/NEI NIH HHS/United States