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The specificity of negative regulation on AR-mediated MMTV-luciferase activity by TR4. Three micrograms of MMTV-Luc was cotransfected with 4 μg of pCMX-TR4 in the presence of 1 μg of pSG5AR, pSG5GR, or pSG5PR by modified calcium-phosphate method. After 24 hr transfection, the cells were treated with 10 nM of synthetic steroids (DHT, dexamethasone, and progesterone). Dual-luciferase reporter assays were performed. pRL-TK was used to normalize the transfection efficiency.
