Rodent chow

China Medical University’s Institutional Animal Care and Treatment Committee approved all protocols involving animals and their welfare. Briefly, 5- to 7-week-old athymic nude mice (female; BALB/c-nu), acquired from the National Laboratory Animal Center (Taipei, Taiwan), were confined in a sterile environment with a 12–12 h light–dark cycle. They were fed rodent chow (Oriental Yeast Co., Tokyo) and provided unlimited access to water.
The mice were subcutaneously engrafted in the right-hind flank with 1 × 10⁶ MDA-MB-231 cells. The mice were separated into two groups of five. For this experiment, cells passaged less than 20 times were used. The treatment-group mice were administered C0Q₀ (0.75 mg/kg b.w.) Intraperitoneal (IP) injection (three times/week) was performed for 12 weeks; the control group received only the vehicle (PBS). The tumors were measured on a weekly basis, and tumor volume was ascertained as width² × length × 0.5 (mm). The mice were sacrificed on the 12th week; this was followed by tumor removal and weighing. A veterinary pathologist examined the excised organs, including the liver, lungs, and kidneys.

In vivo experiments were performed as previously described [70 (link)]. Athymic nude BALB/c-nu mice (8 weeks old) were procured from the National Laboratory Animal Center (NLAC; Taipei, Taiwan). The mice used in this research were kept in a pathogen-free and specially designed animal care facility that offered 12 h of light/dark. The mice used in this research freely accessed water and rodent chow (Oriental Yeast Co Ltd., Tokyo, Japan). This research ensured that all animal protocols were reviewed and approved (approval no: 104-55-N) by Institutional Animal Care and Use Committee (IACUC) of China Medical University. All animal study protocols and methods were conducted following the “Guidelines for the Care and Use of Laboratory Animals”. The aforementioned guidelines were issued by the Chinese Society of Animal Science, Taiwan.

Female athymic nude mice (BALB/c-nu), 5–6 weeks of age, were purchased from The National Laboratory Animal Center (Taipei, Taiwan) and were maintained in caged housing in a specifically designed pathogen-free isolation facility with a 12 h/12 h light/dark cycle. The mice had free access to rodent chow (Oriental Yeast Co Ltd., Tokyo, Japan) and water ad libitum. All animal experiments were strictly followed “The Guidelines for the Care and Use of Laboratory Animals” published by the Chinese Society of Animal Science, Taiwan. The entire animal study protocols were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of China Medical University, Taichung, Taiwan.