24 nude mice (3-4 weeks) were acquired from the experimental animal center of southern medical university (GuangZhou, China) and kept under specific pathogen-free conditions. 4 x 10⁶ PC3 cells were implanted subcutaneously into the right armpit regions of each nude mouse. When the tumors volume reached approximately 40 mm³, mice were randomly divided into two groups to receive flubendazole (10 mg/kg, once daily), 5-fluorouracil (30 mg/kg, once daily), their combination, and vehicle control by intraperitoneal injection. After 20 days of treatment, all mice were sacrificed and tumor weight and tumor volume were immediately measured, respectively.

Adult athymic nude mice were inoculated subcutaneously in the flank with the LNCaP human prostate cancer cell lines in 200 uL of Matrigel. Mice were divided into two groups, and the treatment group was implanted with 2 one cm long silastic implants filled with testosterone as described previously. Tumors were harvested 2- and 4-days post-treatment and fixed in 10% buffered formalin and processed for IHC and H&E staining.

LNCaP cells (5 x 10⁶ cells in 50 uL 10% FBS/RPMI 1640 medium) were co-injected subcutaneously with 50 uL Matrigel in 6-week-old NOD Scid Gamma male mice (Bioresource Facility, Austin Health, Heidelberg, Australia). When tumors reached~200 mm³, mice were randomized in different therapy groups.

20 x 10⁶ cells/mouse were suspended in serum-free medium and mixed with Matrigel (Corning, NY, USA) in a 1:1 ratio. 50 ul of cell suspension were injected orthotopically into the anterior prostate lobe of CD1-nude mice (Charles River Laboratories, Wilmington, MA, USA). Orchidectomy was performed at the time of injection. Tumours were allowed to grow for ~6 weeks after injection and tumour growth was monitored weekly using a Vevo3100 ultrasound imaging system (Fujifilm Visualsonics, The Netherlands).

1 x 10⁶ shCT or shRB PC-3 cells were mixed with 100 uL Matrigel (Corning) and implanted subcutaneously into the right flanks of 6- to 8-week-old male nude mice. When tumor volumes were approximately 80-100 mm³ in PC3 xenografts or circulating RFP tumor cells had begun to emerge in peripheral blood of PPR-RFP mice (around 7.5 months), vehicle or JKE-1674 (25 mg/kg, dissolved in 10% ethanol and 90% PEG-400, Sigma-Aldrich) were administered orally to mice every other day.

A total of 2 x 10⁶ PC3 and PC3/Cd cells were subcutaneously injected into the right flanks of 4-week-old male Balb/c nude mice. Tumor burdens were closely monitored by tumor volumes. When the largest tumors reached a size of 1.0 cm3, all mice were sacrificed due to ethical considerations. Moreover, the final tumor weight was also recorded.

PC3 and PC3/DR cells (5 x 10⁶ cells) were subcutaneously injected into each flank of six-week-old male BALB/c nude mice (HFK Biotech, China). When the tumor volume reached 100 mm³, the mice were treated with Dimethyl Sulfoxide (DMSO) alone, DTX (5 mg/kg body weight, every two days) with DMSO or erastin (20 mg/kg body weight in 20 ul DMSO plus 130 ul corn oil, daily) by intraperitoneal injection.

PC3 and PC3/DR cells (5 x 10⁶ cells) were subcutaneously injected into each flank of six-week-old male BALB/c nude mice (HFK Biotech, China). When the tumor volume reached 100 mm³, the mice were treated with Dimethyl Sulfoxide (DMSO) alone, DTX (5 mg/kg body weight, every two days) with DMSO or erastin (20 mg/kg body weight in 20 ul DMSO plus 130 ul corn oil, daily) by intraperitoneal injection.

PC3 and PC3/DR cells (5 x 10⁶ cells) were subcutaneously injected into each flank of six-week-old male BALB/c nude mice (HFK Biotech, China). When the tumor volume reached 100 mm³, the mice were treated with Dimethyl Sulfoxide (DMSO) alone, DTX (5 mg/kg body weight, every two days) with DMSO or erastin (20 mg/kg body weight in 20 ul DMSO plus 130 ul corn oil, daily) by intraperitoneal injection.

PC3 and PC3/DR cells (5 x 10⁶ cells) were subcutaneously injected into each flank of six-week-old male BALB/c nude mice (HFK Biotech, China). When the tumor volume reached 100 mm³, the mice were treated with Dimethyl Sulfoxide (DMSO) alone, DTX (5 mg/kg body weight, every two days) with DMSO or erastin (20 mg/kg body weight in 20 ul DMSO plus 130 ul corn oil, daily) by intraperitoneal injection.

A total of 2 x 10⁶ PC3 and PC3/Cd cells were subcutaneously injected into the right flanks of 4-week-old male Balb/c nude mice. Tumor burdens were closely monitored by tumor volumes. When the largest tumors reached a size of 1.0 cm3, all mice were sacrificed due to ethical considerations. Moreover, the final tumor weight was also recorded.