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J Immunol Methods. 2015 Aug;423:12-7. doi: 10.1016/j.jim.2015.04.026. Epub 2015 May 9.

Quantification of CD95-induced apoptosis and NF-κB activation at the single cell level.

Author information

1
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Pfälzer Platz, 39106 Magdeburg, Germany. Electronic address: joern.schmidt@med.ovgu.de.
2
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Pfälzer Platz, 39106 Magdeburg, Germany. Electronic address: sabine.pietkiewicz@med.ovgu.de.
3
Institute of Experimental Internal Medicine, Otto von Guericke University, Leipziger Straße 44, 39120 Magdeburg, Germany. Electronic address: naumann@med.ovgu.de.
4
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Pfälzer Platz, 39106 Magdeburg, Germany. Electronic address: inna.lavrik@med.ovgu.de.

Abstract

CD95/Fas/APO-1 is a member of the death receptor (DR) family. Stimulation of CD95 leads to the induction of apoptosis as well as to NF-κB signaling. Crosstalk between these two pathways plays a central role in cell fate. Defects in the regulation of apoptosis and of NF-κB are connected to a number of chronic inflammatory diseases and cancer. For a better understanding of the life/death decisions in the cell and their contribution to disease progression, the development of new technologies is required. Using imaging flow cytometry we developed a method that enables a quantitative detection of different CD95 signaling pathways in the single cell. The important advantage of this method compared to other approaches is that it allows quantifying a large number of single cells undergoing apoptosis and NF-κB activation. This technology could provide new insights into the quantitative characterization of apoptosis and NF-κB at the single cell level and could be used for the quantitative network analysis in systems biology studies.

KEYWORDS:

Apoptosis; CD95/Fas; Imaging flow cytometry; NF-κB; Single cells analysis

PMID:
25967949
DOI:
10.1016/j.jim.2015.04.026
[Indexed for MEDLINE]

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