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Interactions between p65 and Rel-B exist in pre-labour, human amnion.

By Sheri Lim (327707), David A. MacIntyre (230523), Yun S. Lee (327708), Shirin Khanjani (117784), Vasso Terzidou (327709), T. G. Teoh (327710) and Phillip R. Bennett (117789)

Abstract

<p>(<b>A</b>) Whole cell lysate from unstimulated and IL-1β stimulated pre-labour amnion epithelial cells was incubated p65 conjugated beads. The lysate was recaptured under denaturing conditions and Western immunoblotting performed with either anti-p65, anti-phospho-p65 or anti-Rel-B antibodies. Non-stimulated amnion was shown to contain both p65-pp65 and to a greater extent, p65-Rel-B dimers. When stimulated with IL-1β, p65-pp65 dimer levels increased maximally at 30 min and then decreased gradually over 24 h. Dimers of p65-Rel-B were maintained at high levels throughout the time series. (<b>B</b>) Non-radioactive DNA binding assay kit (TRANSAM perbioscience) measuring the binding of Rel-B to the NFκB consensus binding sequence in response to IL-1β showed an increase from the unstimulated state at 30 min before dropping slightly 1 h. Binding peaked at 4 h before again subsiding at 24 h.</p

Topics: Microbiology, Cell Biology, Genetics, p65, rel-b
Year: 2013
DOI identifier: 10.1371/journal.pone.0034707.g003
OAI identifier: oai:figshare.com:article/330169
Provided by: FigShare
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