@article{6f819153a7cd4d74be5d0b9cfc4559fe,
title = "Protocol for electrical modulation of ERK signaling dynamics in cell lines",
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",
author = "Kan Zhu and Liang Guo and Preena Patel and John Albeck and Quan Qing and Wolfgang Losert and Min Zhao",
note = "Funding Information: This work is supported by AFOSR MURI grant ( FA9550-16-1-0052 , Program Leader: Wolfgang Losert, University of Maryland), and also partially supported by DURIP award FA9550-22-1-0149 from the AFOSR, NIH 1R01EY019101 , DARPA grant ( D20AC00003 , Program Leader: Marco Rolando, University of California Santa Cruz), and Adolph Coors Foundation grant. Funding Information: This work is supported by AFOSR MURI grant (FA9550-16-1-0052, Program Leader: Wolfgang Losert, University of Maryland), and also partially supported by DURIP award FA9550-22-1-0149 from the AFOSR, NIH 1R01EY019101, DARPA grant (D20AC00003, Program Leader: Marco Rolando, University of California Santa Cruz), and Adolph Coors Foundation grant. K.Z. L.G. and M.Z. conceived the study. J.A. provided the MCF10A cell line and ERK reporter used in the research. K.Z. and L.G. designed the AC stimulation devices and experimental details. K.Z. and P.P. performed the experiments. K.Z. led the preparation of the manuscript with help from all authors. Q.Q. and W.L. contributed to the improvement of AC devices and data interpretation. M.Z. supervised the study. M.Z. J.A. Q.Q. and L.G. are named inventors with Houpu Li of a U.S. Patent application no. 16/675,127, filed November 5, 2019. Cell signaling pathway activation by local AC electric field. The patent is under consideration. The manuscript used some techniques described in the manuscript to regulate signaling dynamics. Arizona State University and University of California are patent applicants. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = dec,
day = "16",
doi = "10.1016/j.xpro.2022.101752",
language = "English (US)",
volume = "3",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "4",
}