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ASC Speck Formation as a Readout for Inflammasome Activation

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The Inflammasome

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1040))

Abstract

All inflammasomes require the adapter protein apoptosis associated speck-like protein containing a CARD (ASC) for the activation of caspase-1. After inflammasome activation, ASC assembles into a large protein complex, which is termed “speck”. ASC specks can be observed as they reach a size of around 1 μm and in most cells only one speck forms upon inflammasome activation. Hence, ASC speck formation can be used as a simple upstream readout for inflammasome activation. Here, we describe a method for analyzing inflammasome activation by ASC speck visualization. First, we describe the generation of a clonal inflammasome reporter macrophage cell line overexpressing fluorescently tagged ASC. We then discuss stimulation conditions and the microscopic evaluation of ASC speck formation.

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References

  1. Masumoto J, Taniguchi S, Ayukawa K, Sarvotham H, Kishino T, Niikawa N, Hidaka E, Katsuyama T, Higuchi T, Sagara J (1999) ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. J Biol Chem 274(48):33835–33838

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Author information

Authors and Affiliations

  1. Institute of Innate Immunity, University Hospital, University of Bonn, Bonn, Germany

    Andrea Stutz, Gabor L. Horvath, Brian G. Monks & Eicke Latz

  2. Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA

    Brian G. Monks

  3. Division of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA

    Eicke Latz

  4. German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany

    Eicke Latz

Authors
  1. Andrea Stutz
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  2. Gabor L. Horvath
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  3. Brian G. Monks
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  4. Eicke Latz
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Editor information

Editors and Affiliations

  1. , Institute of Innate Immunity, University of Bonn, Sigmund-Freud-Strasse 25, Bonn, 53105, Germany

    Christine M. De Nardo

  2. , Institute of Innate Immunity, University of Bonn, Sigmund-Freud-Str. 25, Bonn, 53105, Germany

    Eicke Latz

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© 2013 Springer Science+business media, New York

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Stutz, A., Horvath, G.L., Monks, B.G., Latz, E. (2013). ASC Speck Formation as a Readout for Inflammasome Activation. In: De Nardo, C., Latz, E. (eds) The Inflammasome. Methods in Molecular Biology, vol 1040. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-523-1_8

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  • DOI: https://doi.org/10.1007/978-1-62703-523-1_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-522-4

  • Online ISBN: 978-1-62703-523-1

  • eBook Packages: Springer Protocols

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