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Wnt/β-catenin TOPFlash assay

QZ Qing Zhong
YZ Yanyu Zhao
FY Fangfei Ye
ZX Zaiyu Xiao
GH Gaoxingyu Huang
MX Meng Xu
YZ Yuanyuan Zhang
XZ Xiechao Zhan
KS Ke Sun
ZW Zhizhi Wang
SC Shanshan Cheng
SF Shan Feng
XZ Xiuxiu Zhao
JZ Jizhong Zhang
PL Peilong Lu
WX Wenqing Xu
QZ Qiang Zhou
DM Dan Ma
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One day before transfection, Hela cells were seeded into 24-well plates. In total, 100 ng (or 300 ng) of BacMam-Wnt3a, 300 ng TOPFlash construct (Beyotime) and 6 ng Renilla luciferase expression construct were co-transfected into Hela cells with 1:2 ratio to lipofectamine 3000. After 36 h’ post-transfection, medium for different Wnt3a variants were collected for Wnt3A secretion detection by western blotting; cells were harvested, and luciferase activities were measured using Dual Luciferase Reporter Gene Assay Kit (Beyotime). Readings for Renilla luciferase activity were used as internal control for Wnt3a activity normalization. Experiments were repeated for at least three times. Full scans of blots and raw data for TOPFlash assay readings are included in the Source Data file.

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