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Luciferase reporter assay

KT Ken-ichi Takayama
TF Tetsuya Fujimura
YS Yutaka Suzuki
SI Satoshi Inoue
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LNCaP and 22Rv1 cells were plated in 24-well dishes at a density of 3 × 104 cells/well in phenol-red-free RPMI medium with 5% charcoal-stripped FBS. After 2 days incubation, the cells were co-transfected with luciferase reporter plasmid (MMTV and PSA-Luc), Renilla luciferase control, and Tk-pRL vector (Promega, Madison, WI) as a reference using X-tremeGene HP DNA Transfection Reagent (Sigma). After 24 h of incubation, the cells were treated with 10 nM DHT or vehicle (0.1% ethanol) for further 24 h. Luciferase activity was measured using the Dual Luciferase Assay Kit (Promega). The firefly luciferase activity was normalized to the Renilla luciferase activity.

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