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Assessing Phosphorylation of STAT Transcription Factors in Mouse Innate Lymphoid Cells

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Innate Lymphoid Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2121))

Abstract

Innate lymphoid cells (ILCs) ensure protection against pathogens by quickly reacting to the alterations of the cytokine milieu taking place upon infection. More than 50 cytokines and growth factors activate the Janus kinases (JAKs), leading to phosphorylation of members of the signal transducer and activator of transcription (STAT) family. Activation of STATs induces specific transcriptional programs which are associated with distinct cellular outcomes. Thus, an efficient measurement of rapid STAT phosphorylation enables not only to dissect the spectrum of cytokine sensitivity among ILC subsets but also to pinpoint specific transcriptional programs and cellular functions initiated after activation. Using this method, we have previously dissected the downstream events of Interleukin (IL)-23 and IL-12 signaling in ILCs, shedding light on the differential usage of STATs among ILC subsets. Here, we provide an optimized and detailed protocol describing how to analyze phosphorylation of STAT transcription factors in murine NK and ILC subsets isolated from different tissues.

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Acknowledgments

G.S. is supported by the Italian Association for Cancer Research (AIRC), MFAG 2018 (Project Code: 21311); and Institut Pasteur (France), Transversal Research Program; PTR-113-17.

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Authors and Affiliations

  1. Department of Molecular Medicine, Sapienza University of Rome, Laboratory Affiliated to Istituto Pasteur Italia—Fondazione Cenci Bolognetti, Rome, Italy

    Gianluca Scarno, Giuseppe Pietropaolo, Chiara Di Censo & Giuseppe Sciumè

  2. Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy

    Giovanna Peruzzi

Authors
  1. Gianluca Scarno
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  2. Giuseppe Pietropaolo
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  3. Chiara Di Censo
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  4. Giovanna Peruzzi
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  5. Giuseppe Sciumè
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Corresponding author

Correspondence to Giuseppe Sciumè .

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Editors and Affiliations

  1. Laboratory of T cell Regulation, Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, UK

    Shoba Amarnath

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Scarno, G., Pietropaolo, G., Di Censo, C., Peruzzi, G., Sciumè, G. (2020). Assessing Phosphorylation of STAT Transcription Factors in Mouse Innate Lymphoid Cells. In: Amarnath, S. (eds) Innate Lymphoid Cells . Methods in Molecular Biology, vol 2121. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0338-3_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0338-3_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0337-6

  • Online ISBN: 978-1-0716-0338-3

  • eBook Packages: Springer Protocols

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