I am going to stimulate fibroblast-like synoviocytes by leptin and TL1A to study the expression of IHH gene. I have wondered, maybe there are special protocols or instructions to follow them in this experiment because I did not try it before.
Depending on what are you measuring you will find "activation" even in the absence of ·activator·. Then run controls without activators and , very important, treat cells as gently as possible.