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2 for 24 h. Ligand stock solutions were prepared (10 mM in DMSO) and diluted 100-fold in media; then, 50 μL of diluted ligand solution was added to each well. The final concentration of DMSO was maintained at 0.03% in both controls and treatments. β-Galactosidase reporter gene expression was measured 48 h after treatment of the cells using Gal Screen bioluminescent reporter gene assay system from Tropix (GSY1000). Luminescence was detected at room temperature using a Dynex MLX microtiter plate luminometer. Fold inductions were calculated by dividing relative light units (RLU) in ligand-treated cells with RLU in DMSO-treated cells.
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