*2.8. Western Blot Analysis*

Extracted proteins were separated by gel electrophoresis and transferred to Immobilon polyvinylidene difluoride membranes (Merck Millipore, Darmstadt, Germany) following our previously reported protocol [27]. The anti-AR, anti-PSA, anti-phospho-Akt (Ser473), anti-Akt, and anti-β-actin antibodies were used at dilutions of 1:2500, 1:5000, 1:1000, 1:1000, and 1:5000, respectively. Specific protein bands were visualized using the SuperSignalTM West Pico Chemiluminescent Substrate (Thermo Fisher Scientific Inc.) with the LAS-4000 Mini (Fuji Photo Film, Tokyo, Japan).
