Luciferase Assay for BV2 Cells

BV2 cells were transfected with 1 μg of reporter plasmid DNA ([κB]₃-luc, ARE-luc, CRE-luc, or PPRE-luc) using Metafectene transfection reagent (Biontex, Martinsried/Planegg, Germany). The effect of GTS-21 on reporter gene activity was assessed by pre-treating the cells with GTS-21 before LPS stimulation (100 ng/mL; 6 h) after 36 h transfection. Following that, cell lysates were collected and a luciferase assay was performed as described previously [29 (link)].

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Park J.E., Leem Y.H., Park J.S., Kim D.Y., Kang J.L, & Kim H.S. (2022). Anti-Inflammatory and Neuroprotective Mechanisms of GTS-21, an α7 Nicotinic Acetylcholine Receptor Agonist, in Neuroinflammation and Parkinson’s Disease Mouse Models. International Journal of Molecular Sciences, 23(8), 4420.

Publication 2022

Metafectene transfection reagent

Assay Cell Gts 21 Luciferase Metafectene Plasmid Reporter gene Transfection

Corresponding Organization : Ewha Womans University

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Variable analysis

independent variables

Transfection of reporter plasmid DNA ([κB]3-luc, ARE-luc, CRE-luc, or PPRE-luc)
Pre-treatment with GTS-21 before LPS stimulation

dependent variables

Reporter gene activity

control variables

Metafectene transfection reagent
LPS stimulation (100 ng/mL; 6 h)
Cell lysate collection and luciferase assay

controls

Positive control: Not specified
Negative control: Not specified

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