Interaction between cultured endothelial cells and macrophages: in vitro model for studying flavonoids in redox-dependent gene expression.
This article focused on two methods to measure the activity of NF-kB. Both methods evalute "post-IkB phosphorylation" stages in the NF-kB activation cascade. In fact, EMSA performed with nuclear extracts provides an information only on NF-kB nuclear translocation and its ability to bind kB-DNA sequences. Likewise, the reporter gene assay is limited to assessing NF-kB-dependent gene expression no matter the mechanism that originally activated NF-kB. Nevertheless, the latter assay represents a more physiological and more reproducible way of measuring NF-kB activity in mammalian cells than the EMSA does. In order to obtain further insights into NF-kB signal transduction pathways, investigating IkB degradation and phosphorylation are recommended. The cloning and characterization of IkB kinases provided new testing possibilities based on measure of their activity.
Rimbach
G
G
School of Food Biosciences, Hugh Sinclair Human Nutrition Unit, University of Reading, Reading RG6 6AP, United Kingdom.
Saliou
C
C
Canali
R
R
Virgili
F
F
D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques; D004727:Endothelium / Q000166:cytology*; D005419:Flavonoids / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017382:Reactive Oxygen Species; D015533:Transcriptional Activation / Q000187:drug effects
D005419:Flavonoids; D016328:NF-kappa B; D017382:Reactive Oxygen Species