Hello, My lab is having great difficulty getting a PPRE luciferase reporter assay to work. I am able to get activation with ligand, but when we do anything else to manipulate the system, we are not getting any induction. This is very strange b/c typically we see coactivation with coactivators, etc. This system has completely stopped working for us. We have gotten new cells, new plasmids, new plasmid preps, new water, new media, you name, it, we've tried it, I think. It has been such a mystery as to why these luciferase assays are no longer showing induction as they were in the past. Any suggestions or ideas would be greatly appreciated.