Proliferation assay

Cells were plated in the appropriate media in quintuplicate (LNCaP: 3,000 cells per well in complete media and 5,000 cells per well in androgen-depleted media; RWPE-1: 2,000 cells per well; RWPE-2: 10,000 cells per well; mouse prostate stem cells: 1,000 cells per well) in poly-d-lysine-coated 96-well plates. Cell proliferation was determined using the Cyquant-NF Cell Proliferation Assay (Invitrogen) or PrestoBlue Cell Viability Assay (ThermoFisher Scientific). Fluorescence was quantified with the SpectraMaxM5 Microplate Reader and SoftMaxPro software (Molecular Devices) and normalized to readings at Day 0 (day after plating).