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HEK293T cells were grown to 70% confluency in 96-well plates (Corning) at 37C in DMEM supplemented with 10% FBS, and treated with Entranster TM -H (Engreen) transfection reagent (0.1 L) and purified plasmids [0.25 g pGL4.74 (hRluc/TK) and 0.25 g pGL4.32(luc2P/NF-B-RE/Hygro plasmid)] in 50 L DMEM/10% FBS per well

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Department of Agricultures recommended daily amount of around 14 grams for every 1,000 calories consumed