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Luciferase reporter assay

XH Xibao Hu
LZ Lei Zhang
JT Jingjing Tian
JM Junhong Ma
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The PART1 wild-type (Wt) and mutant-type (Mut) fragments contained miR-122 binding sites that were synthesized and inserted into pGL3 vector (Promega Corporation, Madison, WI, USA) for getting the PART1-Wt and PART1-Mut reporter plasmid, respectively. Then, 0.5 μg of the reporter plasmids was transfected in combination with 50 pmol of miR-122 inhibitor or inhibitor control by using lipofectamine 2000 reagent according to the manufacturer’s protocol. Luciferase activity was measured 48 h after transfection by using the Dual Luciferase Reporter Assay System (Promega). The relative luciferase activity (Firefly) was normalized to pRL-TK activity (Renilla).

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