Hi everybody, I performed a plasmid transfection on my cancer cells with NDRG-AC-GFP and validated the transfection with both western blotting and IF after 48 and 72 hours. NDRG2 is a transcription factor and it has shown that it effects the protein expression of EMT markers. Unfortunately I couldn't see any effects on EMT marker in my cells. Is it because I am performing a transient transfection? Is 48 and 72 hours enough for a transcription factor to translocate nucleus, bind the promotors and alter both gene and protein expressions? Thanks..