High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells

High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells

Essential characteristics of cellular signaling networks include a complex interconnected architecture and temporal dynamics of protein activity. The latter can be monitored by Förster resonance energy transfer (FRET) biosensors at a single-live-cell level with high temporal resolution. However, the...

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Main Authors: Kuchenov, Dmitry, Laketa, Vibor, Stein, Frank, Salopiata, Florian, Klingmüller, Ursula, Schultz, Carsten
Format: Online
Language:English
Published: Cell Press 2016
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5193178/
id pubmed-5193178
recordtype oai_dc
spelling pubmed-51931782017-01-04 High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells Kuchenov, Dmitry Laketa, Vibor Stein, Frank Salopiata, Florian Klingmüller, Ursula Schultz, Carsten Resource Essential characteristics of cellular signaling networks include a complex interconnected architecture and temporal dynamics of protein activity. The latter can be monitored by Förster resonance energy transfer (FRET) biosensors at a single-live-cell level with high temporal resolution. However, these experiments are typically limited to the use of a couple of FRET biosensors. Here, we describe a FRET-based multi-parameter imaging platform (FMIP) that allows simultaneous high-throughput monitoring of multiple signaling pathways. We apply FMIP to monitor the crosstalk between epidermal growth factor receptor (EGFR) and insulin-like growth factor-1 receptor signaling, signaling perturbations caused by pathophysiologically relevant EGFR mutations, and the effects of a clinically important MEK inhibitor (selumetinib) on the EGFR network. We expect that in the future the platform will be applied to develop comprehensive models of signaling networks and will help to investigate the mechanism of action as well as side effects of therapeutic treatments. Cell Press 2016-12-22 /pmc/articles/PMC5193178/ /pubmed/27939899 http://dx.doi.org/10.1016/j.chembiol.2016.11.008 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Kuchenov, Dmitry
Laketa, Vibor
Stein, Frank
Salopiata, Florian
Klingmüller, Ursula
Schultz, Carsten
spellingShingle Kuchenov, Dmitry
Laketa, Vibor
Stein, Frank
Salopiata, Florian
Klingmüller, Ursula
Schultz, Carsten
High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
author_facet Kuchenov, Dmitry
Laketa, Vibor
Stein, Frank
Salopiata, Florian
Klingmüller, Ursula
Schultz, Carsten
author_sort Kuchenov, Dmitry
title High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
title_short High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
title_full High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
title_fullStr High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
title_full_unstemmed High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells
title_sort high-content imaging platform for profiling intracellular signaling network activity in living cells
description Essential characteristics of cellular signaling networks include a complex interconnected architecture and temporal dynamics of protein activity. The latter can be monitored by Förster resonance energy transfer (FRET) biosensors at a single-live-cell level with high temporal resolution. However, these experiments are typically limited to the use of a couple of FRET biosensors. Here, we describe a FRET-based multi-parameter imaging platform (FMIP) that allows simultaneous high-throughput monitoring of multiple signaling pathways. We apply FMIP to monitor the crosstalk between epidermal growth factor receptor (EGFR) and insulin-like growth factor-1 receptor signaling, signaling perturbations caused by pathophysiologically relevant EGFR mutations, and the effects of a clinically important MEK inhibitor (selumetinib) on the EGFR network. We expect that in the future the platform will be applied to develop comprehensive models of signaling networks and will help to investigate the mechanism of action as well as side effects of therapeutic treatments.
publisher Cell Press
publishDate 2016
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5193178/
_version_ 1613834951827914752

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