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Cell-Based Reporter Assay to Analyze Activation of Nod1 and Nod2

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Immune Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 748))

Abstract

Nod1 and Nod2 are pattern recognition receptors of the mammalian innate immune system. They respond to bacterial peptidoglycan fragments and are implicated in host defense against a variety of ­different bacterial pathogens. Recent studies furthermore support additional functions of these proteins in the control of adaptive immune responses and intestinal homeostasis. Activation of Nod1 and Nod2 by their cognate elicitors triggers inflammatory responses driven by the activation of NF-κB and MAPK pathways. In this chapter, we describe a quick and reliable cell-based assay using a luciferase reporter to measure Nod1- and Nod2-mediated NF-κB activation. The described protocol was successfully applied to analyze the influences of overexpressed proteins and siRNA-mediated knock-down to provide new insights into the regulation of Nod1/2-specific signaling pathways. Furthermore, this method is well suited for downscaling to high-throughput screening applications.

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Acknowledgments

We thank Dr. Katja Lautz and Andreas Neerincx for critical reading of the manuscript. The authors acknowledge support by the Deutsche Forschungsgemeinschaft (DFG) grant SFB670-NG01.

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Authors and Affiliations

  1. Molecular Innate Immunobiology Group, Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany

    Thomas A. Kufer

Authors
  1. Birte Zurek
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  2. Harald Bielig
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  3. Thomas A. Kufer
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Corresponding author

Correspondence to Thomas A. Kufer .

Editor information

Editors and Affiliations

  1. Sunnybrook Health Sciences Centre, University of Toronto, Bayview Avenue 2075, Toronto, M4N 3M5, Ontario, Canada

    Jonathan P. Rast

  2. Sunnybrook Research Institute, Department of Immunology, University of Toronto, Bayview Avenue 2075, Toronto, M4N 3M5, Ontario, Canada

    James W.D. Booth

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Zurek, B., Bielig, H., Kufer, T.A. (2011). Cell-Based Reporter Assay to Analyze Activation of Nod1 and Nod2. In: Rast, J., Booth, J. (eds) Immune Receptors. Methods in Molecular Biology, vol 748. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-139-0_7

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  • DOI: https://doi.org/10.1007/978-1-61779-139-0_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-138-3

  • Online ISBN: 978-1-61779-139-0

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