Dual-Luciferase Reporter Assay for AP-1 Activity

The Dual-Luciferase reporter assay was performed as described previously [28 (link)]. Briefly, the ESCC cells were seeded in a 24-well plate and transfected with pGL3-AP-1 (#40342, Addgene) or pGL3-Basic control vector along with the renilla luciferase reporter construct pRL-SV40for 24 h. Cells were then treated with isoliquiritigenin for another 24 h and harvested for firefly luciferase and renilla luciferase activity determination using the Dual-Luciferase reporter assay system (#E1910; Promega, Madison, WI, USA). Renilla luciferase activity was used as a control for transfection efficiency.

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Ye L., Zhang J., Zhang Y., Gu B., Zhu H, & Mao X. (2020). Isoliquiritigenin Suppressed Esophageal Squamous Carcinoma Growth by Blocking EGFR Activation and Inducing Cell Cycle Arrest. BioMed Research International, 2020, 9259852.

Publication 2020

pGL3-AP-1 Dual-Luciferase reporter assay system

Assay Cells Firefly luciferase Isoliquiritigenin Luciferase Pgl3 Promega Renilla luciferase Transfection

Corresponding Organization : Wenzhou Medical University

Other organizations : Tongji University, Tongji Hospital

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Variable analysis

independent variables

Isoliquiritigenin

dependent variables

Firefly luciferase activity
Renilla luciferase activity

control variables

Transfection efficiency (measured by Renilla luciferase activity)

controls

Positive control: pGL3-AP-1 reporter construct
Negative control: pGL3-Basic control vector

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