Luciferase Assay for NF-κB Activity

The experiment was performed as described [23 (link)]. NF-κB promoter region was cloned into pGL3-based vectors to construct Luciferase reporter plasmid. The reporter plasmid was transfected into RA-FLS-Ctrl or RA-FLS-TRIP groups using the Lipofectamine 2000 (Invitrogen, Shanghai, China), and phRL-TK plasmid was cotransfected as internal control. 36 hours later, the Luciferase activities were measured on a SpectraMax M5 reader (Molecular Devices, California, USA) using the Dual-Luciferase Reporter Assay System (Promega, Madison, USA).

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Kong Q.Z., Guo L.T., Yang J.N., Wang Y.F., Zhao J.X., Kong S.H., Zhang M., Yan S, & Jin Y. (2016). Anti-Inflammatory Effects of TRAF-Interacting Protein in Rheumatoid Arthritis Fibroblast-Like Synoviocytes. Mediators of Inflammation, 2016, 3906108.

Publication 2016

Lipofectamine 2000 SpectraMax M5 reader Dual-Luciferase Reporter Assay System

Assay Devices Lipofectamine 2000 Luciferase Nf κb Pgl3 Plasmid Promega Vectors

Corresponding Organization : Chengde Medical University

Other organizations : Maternal and Child Health Hospital of Sichuan Province

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Variable analysis

independent variables

Experimental group (RA-FLS-Ctrl or RA-FLS-TRIP)

dependent variables

Luciferase activity

control variables

Transfection efficiency (phRL-TK plasmid as internal control)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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