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Dual-luciferase reporter assay

RZ Ruinan Zhao
XG Xiangyu Guo
GZ Guohao Zhang
SL Sen Liu
RM Ranran Ma
MW Mengqi Wang
SC Shiming Chen
WZ Wenjie Zhu
YL Yuan Liu
PG Peng Gao
HL Haiting Liu
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Following the manufacturer’s instructions, a dual-luciferase reporter assay (Promega) was used to detect the firefly luciferase and renilla luciferase signals. Briefly, MKN-45 and BGC-823 cells were seeded overnight in 24-well plates and co-transfected with Lipofectamine 2000 (Invitrogen) using 0.5 μg of the overexpression vector, 0.5 μg of the HNF1A-AS1 promoter-luciferase reporter plasmid, and 0.01 μg of pRL-TK plasmid. Internal control was implemented using PRL-TK. After approximately 48 h of transfection, the luciferase activity of the cells was determined using a Dual-Luciferase Reporter Assay System (Promega) in accordance with the manufacturer’s instructions. Relative fluorescence values were expressed as firefly fluorescence values over renilla fluorescence values.

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