2.1. Enhanced Green Fluorescent Protein (eGFP) Cellular AR Transcription Assay

AR transcriptional activity was assayed as previously described [55]. Briefly, LNCaP human prostate cancer cells (LN-ARR2PB-eGFP), stably transfected with eGFP reporter driven by an androgen responsive probasin-derived promoter (ARR2PB), were grown in phenol red free RPMI 1640 media supplemented with 5% charcoal stripped serum (CSS). After 5 days, the cells were plated into a 96-well plate (35,000 cells/well) with 0.1 nM of the synthetic androgen R1881 and increasing concentrations (0−100 μM) of test compounds. Fluorescence (excitation 485 nm, emission 535 nm) was measured after 3 days of incubation with the compounds.