Cell proliferation assay

LNCaP cells or C4–2B cells (750 cells/well) were seeded in 96-well plates. After 16 hours of incubation, cells were treated with ethanol (control) or the indicated doses of LLM or synthetic androgen R1881 for 24, 48, and 72 hours. Subsequently, 20 μL of manufacturer supplied colour development reagent (MTS, Promega, Madison, WI) was added to each well and the plates were incubated at 37°C for 2 hours. Absorbance was measured at 492 nm.