Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells


Journal article


I. Weibrecht, I. Grundberg, M. Nilsson, O. Söderberg
PLoS ONE, 2011

Semantic Scholar DOI PubMedCentral PubMed
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APA   Click to copy
Weibrecht, I., Grundberg, I., Nilsson, M., & Söderberg, O. (2011). Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells. PLoS ONE.


Chicago/Turabian   Click to copy
Weibrecht, I., I. Grundberg, M. Nilsson, and O. Söderberg. “Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells.” PLoS ONE (2011).


MLA   Click to copy
Weibrecht, I., et al. “Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells.” PLoS ONE, 2011.


BibTeX   Click to copy

@article{i2011a,
  title = {Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells},
  year = {2011},
  journal = {PLoS ONE},
  author = {Weibrecht, I. and Grundberg, I. and Nilsson, M. and Söderberg, O.}
}

Abstract

We have developed an approach for simultaneous detection of individual endogenous protein modifications and mRNA molecules in single cells in situ. For this purpose we combined two methods previously developed in our lab: in situ proximity ligation assay for the detection of individual protein interactions and -modifications and in situ detection of single mRNA molecules using padlock probes. As proof-of-principle, we demonstrated the utility of the method for simultaneous detection of phosphorylated PDGFRβ and DUSP6/MKP-3 mRNA molecules in individual human fibroblasts upon PDGF-BB stimulation. Further we applied drugs disrupting the PDGFRβ signaling pathway at various sites to show that this combined method can concurrently monitor the molecular effect of the drugs, i.e. inhibition of downstream signaling from the targeted node in the signaling pathway. Due to its ability to detect different types of molecules in single cells in situ the method presented here can contribute to a deeper understanding of cell-to-cell variations and can be applied to e.g. pinpoint effector sites of drugs in a signaling pathway.


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