Hello everyone.
I have generated DU145- NF-kB reporter cell line (luciferase). In these cells, NF-kB are getting activated with TNF, but not getting inhibited with curcumin or other known NF-kB inhibitor. Please suggest possible reasons.
Hello,
You can check different kinase activities whether they are influencing on curcumin effects. You also should check whether you transfected the cells properly for greater expressions of NF-kB.
Thank you for your reply. I have generated these cells with lenti virus and selected with puromycin. Basal activity is detectable.
First do a viability assay or kill curve analysis to find out what dosages of the inhibitor are not lethal to the cell line. Second, pre-treat the cells first with inhibitor for 30 min-4h (or more), you will need to try various exposure timings. After pre-treatment you should proceed to add the activator and then check for inhibition.
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