I'm doing a preliminary experiment to induce CXCL9 via TNFa in Endothelial cells. The plan for the experiment is to bring up some Endothelial cells and expose them to TNFa (as we already have this in stock else we would use IFN-y). Has anyone got any experience in what concentration of TNFa and the duration you would use to start this experiment or could point me to the literature that might be useful?
Many thanks for reading.
Dear Calum,
20 ng/ml TNF-a typically result in a strong activation of the NF-kB pathway in endothelial cells and activation of target genes. Depending on the gene, you may see the strongest induction after 3-6 h but in some cases also later, e.g. after 24 h. However, in case of CXCL9, I doubt that endothelial cells will upregulate this gene very much. If at all, you will probably see a better response using IFN-g (100 ng/ml) or a combination of TNF-a and IFN-g.
Best, Lothar
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