*5.2. Chemicals*

NCL1 was synthesized as previously described [17].

### *5.3. Prostate Cancer Cell Lines*

The human prostate cancer cell lines PC3 and 22Rv1 were obtained from the American Type Culture Collection (Rockville, MD, USA); these cells plus a castration-resistant rat prostate cancer cell line, PCai1, were cultured as previously described [46,47]. The cell lines used included PC3, which is human CRPC cell line without AR expression; 22Rv1, which is human CRPC cell line with AR expression; and PCai1CS, which is established from CRPC originating from a transgenic rat model as previously described [48,49]. They were cultured in media with 10% charcoal-stripped serum, and then treated with dimethyl sulfoxide (DMSO) as a vehicle that was equal in concentration to that used for 1–100 μM NCL1 for 72 h. Finally, its effect on cell proliferation determined. To assess the effects of autophagy on cell proliferation, PC3 and 22Rv1 cells were treated with 50 μM NCL1 and/or 50 μM chloroquine (CQ) for 24 h, an inhibitor of autophagy. All experiments were performed in triplicate.
