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Transfection and luciferase reporter gene assays
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Cells plated on 24-well dishes were transfected using Lipofectamine 3000 (Invitrogen) for 24 h, after which they were serum-starved for an additional 24 h prior to harvest. A renilla luciferase reporter vector pRL-TK was used as the internal control of transfection efficiency. Cells were subjected to luciferase activity assays using a dual-specific luciferase assay kit (Promega, Madison, WI, USA). Firefly luciferase activities were normalized on the basis of renilla luciferase activities.
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