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To analyze ERE promoter activity, HCC1806 cells stably expressing the indicated shRNA were co-transfected with the ERE-containing luciferase reporter plasmid or identical NON-Luc construct plus pRL-TK-Luc to assess transfection efficiency, and then treated with 10nM E2 for 24h. Cells were harvested and the luciferase activities were measured using Promega's Luciferase Assay System (Promega, Madison, Wisconsin). The luciferase values (relative light units, RLUs) were normalized based on the activity of pRL-TK-Luc.
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