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Neutrophil pp 261-275 | Cite as

Detection of Intact Transcription Factors in Human Neutrophils

  • Patrick P. McDonald
  • Richard D. YeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2087)

Abstract

The crucial contribution of neutrophils to innate immunity extends well beyond their traditional role as professional phagocytes. Indeed, it is now well established that neutrophils generate a plethora of inflammatory cytokines and chemokines that are profoundly involved in the onset and evolution of the inflammatory reaction. Several recent studies have shown that neutrophils can represent an important source of inflammatory cytokines in pathophysiological settings. The inflammatory and immunomodulatory cytokines produced by neutrophils are generally encoded by immediate-early response genes, which in turn depend on the activation of transcription factors such as those belonging to the nuclear factor κB (NF-κB) and signal transducers and activators of transcription (STAT) families. We have shown in the past that the expression of such factors is induced in neutrophils stimulated by physiological agonists. However, the detection of intact (i.e., undegraded) transcription factors in neutrophils requires special precautions and a specially designed protocol, due to the huge amounts of endogenous proteases present in these cells. This protocol is the focus of this chapter.

Key words

Transcription factors NF-κB STAT Nuclear extracts Electrophoresis mobility shift assay Granulocytes 

Notes

Acknowledgments

This work was supported by grants to PPMcD from the Canadian Institutes for Health Research and the Arthritis Foundation of Canada, and to RDY from the National Institutes of Health. PPMcD is a scholar of Fonds de la recherche en santé du Québec (FRSQ).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Pulmonary Division, Medicine FacultyUniversité de SherbrookeSherbrookeCanada
  2. 2.Kobilka Institute of Innovative Drug Discovery, School of Life and Health SciencesThe Chinese University of Hong KongShenzhenChina

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