Journal article
Establishment of a T‐Ag and NF‐κB binding elements gene co‐transfected stable HUVECs cell line
Journal of cellular biochemistry, Vol.89(6), pp.1285-1290
08/15/2003
DOI: 10.1002/jcb.10588
PMID: 12898524
Abstract
To set up a T‐Ag gene transfected stable human umbilical venous endothelial cells (HUVECs) cell line and a T‐Ag, nuclear transcriptional factor kappa B (NF‐κB) binding elements linked with luciferase reporter gene co‐transfected stable HUVECs cell line. Cultured HUVECs were transfected with pCI‐neo‐T‐Ag and pRSV‐luc‐3XκB by lipofectin. The G418 selected monoclones were subcultured. The expression of marker protein, vWF and the characteristic of uptake of lipids were compared by Western blotting and immunocytochemistry in non‐transfected and transfected HUVECs. The reporter gene assay was done in the presence of TNF‐α. A T‐Ag gene transfected stable HUVECs cell line and a T‐Ag and NF‐κB binding elements linked with luciferase reporter gene co‐transfected stable HUVECs cell lines were set up. The expression of vWF of these cell lines was similar with those in non‐transformed HUVECs. The function of uptaking of lipids was preserved as well in transfected cell lines. Furthermore, TNF‐α, a typical cytokine increasing the activity of NF‐κB was used to treat the transfected cells O/N. The higher luciferase reporter gene activity was seen. A pCI‐neo‐T‐Ag and pRSV‐luc‐3X κB co‐transfected stable HUVECs cell line might be used to check reporter gene activity directly. It might be a useful tool to screen drugs acting on transcription level. © 2003 Wiley‐Liss, Inc.
Details
- Title: Subtitle
- Establishment of a T‐Ag and NF‐κB binding elements gene co‐transfected stable HUVECs cell line
- Creators
- Y.X LiuW.S CaoK IraniX.J Li
- Resource Type
- Journal article
- Publication Details
- Journal of cellular biochemistry, Vol.89(6), pp.1285-1290
- Publisher
- Wiley Subscription Services, Inc., A Wiley Company; Hoboken
- DOI
- 10.1002/jcb.10588
- PMID
- 12898524
- ISSN
- 0730-2312
- eISSN
- 1097-4644
- Number of pages
- 6
- Language
- English
- Date published
- 08/15/2003
- Academic Unit
- Cardiovascular Medicine; Radiation Oncology; Fraternal Order of Eagles Diabetes Research Center; Internal Medicine
- Record Identifier
- 9984047880302771
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