I have a plan to test the effect of phenolic compounds on LPS-treated cells and show its effect on pathways, because of that I am going to use pathways inhibitors. How to treat cells with LPS, Phenolic compound and inhibitor at the same time?
Assuming you can get all of these things into solution at a sufficient concentration to test them without killing the cells, the problem will be to distinguish the effects of each substance. If you are interested in LPS-treated cells only, then there are 2 variables: concentration of phenolic compound x concentration of inhibitor.
You should vary the concentration of phenolic compound in one dimension and vary the concentration of inhibitor in the other dimension. Then you can measure the effect on the outcome of interest as a function of each combination. The result can be used to determine of there is synergy or antagonism between the two treatments using the fractional inhibitory concentration index (FICI) "checkerboard" method, which is used for testing combinations of antibiotics.
Article The fractional inhibitory concentration (FIC) index as a mea...
Repeat the experiment with and without a fixed concentration of LPS to determine whether the FICI is affected by LPS.
The problem is in the order of treatment. Is LPS treatment prior to inhibitor and phenolic compound or vice versa?
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