Detection of Intact Transcription Factors in Human Neutrophils.
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.
['McDonald|Patrick P|PP|', 'Ye|Richard D|RD|']
D001681:Biological Assay D015415:Biomarkers D002467:Cell Nucleus D024202:Electrophoretic Mobility Shift Assay D006801:Humans D016328:NF-kappa B D009504:Neutrophils D050791:STAT Transcription Factors D014157:Transcription Factors
Transcription factors NF-κB STAT Electrophoresis mobility shift assay Granulocytes Nuclear extracts