Hey guys, I got headache problem with my miRNA functional study. I transfected with the miRNA, and then did a lot of western blot. and then i found all the proteins I tested were up-regulated by the miRNA, including those apparently act opposite roles (e.g. pro-apoptosis vs. anti-apoptosis; cell cycle arrest vs. proliferation related protein) and those predicted as target proteins! even more surprisingly, there was one predicted target which showed promising results in the 3'UTR-luciferase assay, again appeared up-regulated in the western blot. I am sure I applied the samples in the right order in all the assay. Anyone advise why it is happening?! Thanks!
problem with miRNA functional study
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