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Luciferase reporter assay to assess Wnt/β-catenin activity

MP Mineon Park
YC Yong Jin Cho
BK Bora Kim
YK Young Jong Ko
YJ Yuria Jang
YM Yeon Hee Moon
HH Hoon Hyun
WL Wonbong Lim
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Cells were seeded into a 24-well plate 24 h prior to transient transfection with either 2 μg of TOP flash or FOP flash reporter plasmid along with 1 μg DNA using Lipofectamine 3000 (Thermo Fisher). The TOPflash luciferase reporter plasmid contains TCF-4-binding sites upstream of the luciferase gene, resulting in luciferase activity in the presence of active Wnt/β-catenin signalling. The FOP flash reporter plasmid, on the other hand, carried mutated TCF-4-binding sites. Total cell extracts were assayed for luciferase activity according to the manufacturer’s instructions (Promega).

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