Doxycycline free foetal bovine serum

Cells were maintained at 37 °C in 5% CO_2. LNCaPs were maintained in RPMI medium with 10% foetal bovine serum (First Link, Wolverhampton, UK) (ATCC CRL-1740). LNCaP/PHB^cDNA and LNCaP/PHB^siRNA cells^{5 (link)} were maintained in RPMI medium with 10% doxycycline-free foetal bovine serum (Clontech, Palo Alto, CA, USA), 12 μg/ml blasticidin (Sigma, Dorset, UK), 0.3 mg/ml zeocin (ThermoFisher, Waltham, MA, USA) and 500 μg/ml G418 (Sigma). COS-7 cells (ECACC 87021302), PC3 cells (ATCC CRL-1435) and VCaP (ATCC CRL-2876) were maintained in Dulbecco’s modified Eagle's medium with 10% foetal bovine serum. Media were supplemented with 2 mMl-glutamine, 100 U/ml penicillin and 100 mg/ml streptomycin (Sigma). Seventy-two hours before androgen exposure, the medium was replaced with the ‘starvation medium’ consisting of phenol red-free RPMI medium (or Dulbecco’s modified Eagle's medium), supplemented with 5% charcoal-stripped foetal bovine serum (First Link).

Cells were maintained at 37 °C in 5% CO₂. LNCaP cells were maintained in RPMI medium with 10% foetal bovine serum (First Link, Wolverhampton, UK) (ATCC CRL-1740). LNCaP/Luc/PHB^cDNA and LNCaP/Luc/PHB^siRNA cells were maintained in RPMI medium with 10% doxycycline-free foetal bovine serum (Clontech, Palo Alto, CA, USA), 12 μg/mL blasticidin (Sigma, Dorset, UK), 0.3 mg/mL Zeocin (ThermoFisher, Waltham, MA, USA) and 500 μg/mL G418 (Sigma). HeLa (ATCC CCL-2) and PC3 (ATCC CRL-1435) were maintained in Dulbecco’s modified Eagle’s medium with 10% foetal bovine serum. Media were supplemented with 2 mM L-glutamine, 100 U/mL penicillin and 100 mg/mL streptomycin (Sigma). Seventy-two hours prior to androgen exposure, the medium was replaced with the ‘starvation medium’ consisting of phenol-red-free RPMI medium (or Dulbecco’s modified Eagle’s medium), supplemented with 5% charcoal-stripped foetal bovine serum (First Link).