Please tell which LPS (strain of E.coli) would give best results. Time of induction? what about starvation condition ?
I tried with 100ng/ml to 4 ug/ml but could'nt get COX 2 or cCaspase 1?
See this paper here, on HepG2. Perhaps you need to check the serum levels you used.
http://www.nature.com/pr/journal/v45/n4-2/full/pr19991145a.html
Also, supplements like glutamine, and pyruvate may also contribute to a blunted effect.
Some cancer lines have been reported to be frequently infected, even when directly obtained from ATCC. You may want to check your cell line's microbial background if any. A line infected with mycoplasma may not behave quite predictably as you may want it to.
Good luck
Did u checked the levels of IL6, IL8 and TNF-alpha. What is the expression of TLR4 receptor in gastric cell lines u used. Generally LPS at 10ng/ml to 1ug/ml within 24hours induces IL6, IL8 and TNF-alpha by many folds. As Wong mentioned above check cells for mycoplasma contamination.
In order to activate caspase-1, two signals are usually needed: first, LPS stimualtion to induce expression of the inflammasome components, and second, signal 2 such as ATP to activate NLRP3 inflammasome leadind to caspase-1 cleavage (activation).
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