Abstract
Understanding how signaling networks are regulated offers valuable insights into how cells and organisms react to internal and external stimuli and is crucial for developing novel strategies to treat diseases. To achieve this, it is necessary to delineate the intricate interactions between the nodes in the network, which can be accomplished by measuring the activities of individual nodes under perturbation conditions. To facilitate this, we have recently developed a biosensor barcoding technique that enables massively multiplexed tracking of numerous signaling activities in live cells using genetically encoded fluorescent biosensors. In this chapter, we detail how we employed this method to reconstruct the EGFR signaling network by systematically monitoring the activities of individual nodes under perturbations.
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Acknowledgments
The plasmids of PicchuEV, EV-ROCK, and EV-S6K were provided by Michiyuki Matsuda, to whom the authors extend their gratitude. This work was supported by funding from R01GM136711 (to C.H.H.), Cervical Cancer SPORE P50CA098252 Career Development Award (to J.M.Y.) and Pilot Project Award (to C.H.H.), the W. W. Smith Charitable Trust (#C1901, to C.H.H.) and the Sol Goldman Pancreatic Cancer Research Center (to C.H.H.). Additionally, the purchase of the Zeiss LSM 780 confocal microscope was made possible by NIH grants S10OD016374.
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Wang, S., Chi, WY., Au, G., Huang, CC., Yang, JM., Huang, CH. (2024). Reconstructing Signaling Networks Using Biosensor Barcoding. In: Wuelfing, C., Murphy, R.F. (eds) Imaging Cell Signaling. Methods in Molecular Biology, vol 2800. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3834-7_13
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DOI: https://doi.org/10.1007/978-1-0716-3834-7_13
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