C57bl 6 male mice

The present study used a subcutaneous model involving male BALB/c nude mice 4–6 weeks old for investigating tumour growth. Specifically, 5 × 10⁶ transfected cells were suspended in 100 μl of PBS and subcutaneously injected into the right mid‐posterior axilla of the nude mice. Tumour volumes were measured on a weekly basis, and after 5 weeks the mice were euthanized and the tumour weights were recorded. Subsequently, subcutaneous tumours were collected for further analysis via IF and IHC staining. To establish a lung metastases model, 1 × 10⁶ transfected cells were injected into the tail vein of each mouse. The lung tissues from each mouse were collected and stained with H&E after 60 days of tumour development. Pulmonary metastases were counted and survival curves were plotted.
The experimental use of transgenic TRAMP+ mice, which were derived from C57BL/6 male mice obtained from the Jackson Laboratory. Prostatic tissue samples were collected from two age groups (12 weeks [n = 5 per group] and 37 weeks [n = 5 per group]) and classified by pathologists based on histologic features as normal tissues, high‐grade PIN (HGPIN) and UD‐adeno.

Seven-week-old C57BL/6 male mice (n = 18) (Jackson Laboratories, Bar Harbor, ME, USA) were group housed on a reverse light cycle for at least one week prior to the experiment. HA mice were group housed in conventional cages inside a modified Vicker’s hypobaric chamber (Reimers System Inc., Lorton, VA, USA) for 12 weeks to operate under reduced pressures (~7.4 psi) using a vacuum pump (Welch Model 2585B or 2067B-01, Mount Prospect, IL, USA) as previously described^{13 (link),19 (link)}. To avoid acute effect, ascent to a simulated altitude of 5000 m (HA) proceeded at 200 m per minute and then returned to SL at the same rate. Environmental parameters (pressure, oxygen, carbon dioxide levels, temperature and relative humidity) inside of the chamber were continuously monitored using custom built sensors from CO₂ Meter Inc. (Ormond Beach, FL, USA). The chamber was located within the animal vivarium to ensure consistent environmental parameters across exposure groups and conditions. The chamber altitude was monitored using a data logging digital manometer (AZ Instrument Corp., Taichung City, Taiwan). Cage maintenance was performed at SL at least once per week. All mice were monitored daily for signs of distress including failure to groom, and/or excessive weight loss.