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TOP/FOP Flash Luciferase Assay

XW Xi Wang
BL Bin Lu
CD Chunyan Dai
YF Yufei Fu
KH Ke Hao
BZ Bing Zhao
ZC Zhe Chen
LF Li Fu
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TOP/FOP flash luciferase assay was used to detect the transcriptional activity of the TCF-dependent WNT/β-catenin signaling. The reporter plasmid of β-catenin (TOP Flash) and its mutant control (FOP Flash) were from Dr. Fu (Shenzhen University, China). AGS cells were grown in a 24-well plate (5 × 104 cells per well) overnight and transiently transfected with Cav-1 overexpression vector or empty vector for 24 h. Subsequently, the cells were co-transfected with TOP or FOP flash luciferase plasmids and pRL-TK vector (also from Dr Fu) for 24 h. The activity of TOP/FOP flash reporter was examined by Promega Dual-Luciferase Reporter Assay System, and the β-catenin-driven transcription was presented as the TOP/FOP ratio which was normalized to the relative ratio of Renilla Luciferase activity, respectively.

Copyright and License information:  ©2020 Wang, Lu, Dai, Fu, Hao, Zhao, Chen and Fu.
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