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Examining the Kinetics of Phagocytosis-Coupled Inflammasome Activation in Murine Bone Marrow-Derived Dendritic Cells

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Phagocytosis and Phagosomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2692))

Abstract

In the present chapter, we describe procedures to assess NLRP3 and NLRC4 inflammasome assembly by immunofluorescence microscopy or live cell imaging, together with inflammasome activation by biochemical and immunological techniques upon phagocytosis. We also include a step-by-step guide to automating the counting of inflammasome “specks” after imaging. While our focus resides on murine bone marrow-derived dendritic cells differentiated in the presence of granulocyte-macrophage colony-stimulating factor, which results in a cell population that resembles inflammatory dendritic cells, the strategies described herein may apply to other phagocytes as well.

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Acknowledgments

This work was supported by NIH grant R01 AI137173. The summary figure was created using Biorender.com through Thomas Jefferson University library portal.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA, USA

    Daniel J. Netting & Adriana R. Mantegazza

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  1. Daniel J. Netting
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  2. Adriana R. Mantegazza
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Correspondence to Adriana R. Mantegazza .

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  1. Department of Chemistry and Biology, Toronto Metropolitan University, Toronto, ON, Canada

    Roberto J. Botelho

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Netting, D.J., Mantegazza, A.R. (2023). Examining the Kinetics of Phagocytosis-Coupled Inflammasome Activation in Murine Bone Marrow-Derived Dendritic Cells. In: Botelho, R.J. (eds) Phagocytosis and Phagosomes. Methods in Molecular Biology, vol 2692. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3338-0_20

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  • DOI: https://doi.org/10.1007/978-1-0716-3338-0_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3337-3

  • Online ISBN: 978-1-0716-3338-0

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