*4.6. TEM*

The ventral prostates were washed with normal saline solution and cut into pieces of about 1 mm<sup>3</sup> on ice, and fixed with a 2.5% glutaraldehyde solution for 2 h at 4 ◦C. Following three washes with 0.1 M PBS, the tissues were postfixed in 1% osmium tetroxide for 2 h. After dehydration in ascending concentrations of ethanol and acetone, the tissues were embedded in Epon 618. Thin 70 nm sections stained with lead citrate were examined under a transmission electron microscope.

#### *4.7. Analysis of SOD Activity*

The frozen prostate tissue was homogenized in cold normal saline solution. The homogenate was centrifuged at 1520× *g* for 10 min at 4 ◦C. Then, the supernatants were collected, and the Bradford protein assay (Bradford assay) was used to quantify the protein amount. The SOD activity was assessed strictly according to the manufacturer's protocol, as previously reported [32–34].

#### *4.8. Analysis of Levels of T and DHT*

The frozen prostate tissue was homogenized in cold normal saline solution. After two freeze-thaw cycles, the homogenate was centrifuged at 5000× *g* for 5 min at 4 ◦C. The supernatants were collected, while protein amounts were quantified by Bradford assay. Levels of T and DHT were assessed strictly according to the manufacturer's protocol, as previously reported [35–37].
