Firefly Luciferase Reporter Assay Protocol

The protocol for the firefly luciferase reporter assay was described previously [18 (link)]. Briefly, 3.0 g of TCF/LEF luciferase reporter was added to culture flasks, and transfection was performed with Lipofectamine^TM 2000 (Invitrogen, Gaithersburg, MD, USA). Twenty-four hours after transfection, the treated cells were replanted in 24-well plates, and EGCG or DMSO was then added to the cells in each group. After 24 hours, the upper medium was collected for fluorescence activity detection using a Gaussia Luciferase Assay kit (New England Biolabs).

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Dong C., Wang Z., Shen P., Chen Y., Wang J, & Wang H. (2022). Epigallocatechin-3-gallate suppresses the growth of human osteosarcoma by inhibiting the Wnt/β-catenin signalling pathway. Bioengineered, 13(4), 8490-8502.

Publication 2022

LipofectamineTM 2000 Gaussia Luciferase Assay kit

Assay Cells Dmso Egcg Firefly luciferase Fluorescence Luciferase Tcf 3 Transfection

Corresponding Organization : First People's Hospital of Chongqing

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Variable analysis

dependent variables

Luciferase reporter activity

control variables

TCF/LEF luciferase reporter
Lipofectamine2000 for transfection
Cell culture conditions (e.g., cell type, incubation time)

controls

Positive control: Not specified
Negative control: DMSO

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