SBF-1/DHT competitive assay
Flag-ARWT construct was transfected into PC3 cells and the total protein extract was obtained and purified along with Flag tag removal using enterokinase enzyme as reported (Skala, Goettig & Brandstetter, 2013). SBF-1 was checked whether being binding to the ligand binding domain of AR (AR-LBD) or not by a competitive assay using substrate Fluormone AL Green (Thermo Fisher) according to manufacture protocol.