Nuj foxn1nu

All mice studies were approved by the Stony Brook University Institutional Animal Care and Use Committee (IACUC protocol number: 354445) and performed in accordance with institutional policies,NIH and ARRIVE guidelines. Nude mice were purchased from Jackson Laboratories (NuJ/Foxn1^nu, Stock number: 002019, Bar Harbor,). Animals were housed under specific pathogen-free conditions in ventilated and filtered cages under positive pressure. DLD-1 human colorectal cells at concentration of 5×10⁶ were subcutaneously injected into the right flank of 7 week old male nude mice. Tumor volume was determined by caliper measurement and calculated by the established method (25 (link)). When tumors reached a volume of about 100 mm³, mice were treated intraperitoneally (i.p.) with varying doses of SR18662: 5mg/kg daily, 5mg/kg twice a day,10 mg/kg daily, 10 mg/kg twice per day, 25mg/kg daily, and 25 mg/kg twice per day. Each treatment regimen was as follows: 5 days of injections, 2 days break, and 5 days of injections, and mice were collected 24 h after the last injection. The vehicle solution (30% 2-hydroxypropyl-beta-cyclodextrin) was used as the control treatment. Mice were monitored and weighed every two days. Tumors were excised and retained for further analyses.

Female homozygous nude mice (Nu/J Foxn1 nu) at the age of 5 weeks were purchased from the Jackson Laboratory and quarantined for one week in Ohio University animal facility before tumor inoculation. A375 cells were prepared in PBS and mixed with Matrigel (Corning) at equal volume, 100 μL of the mixture (5x10⁵ cells) was injected into mouse subcutaneously. After injection, mice were randomly divided into control and treatment groups. Control vehicle or CaeA (5 mg/kg) was administrated via intraperitoneal injection three times a week starting from the second day of tumor injection. When tumors became palpable, the concentration of CaeA was increased to 10 mg/kg. Approximately 4 weeks after tumor injection, mice were euthanized, and tumors were surgically removed for comparison. All the experimental protocols with animals were approved by the Institutional Animal Care & Use Committee of Ohio University (Protocol Number: 19-H-015).

All animal studies were performed according to procedures approved by the University Committee on Use and Care of Animals. NU/J Foxn1^nu (nude) and C57BL/6 wild type mice were obtained from the Jackson Laboratory. Mice were maintained under 12/12hr light/dark cycle and fed with standard rodent chow. Mice were injected intraperitoneally daily with saline or CL316,243 (1 mg/kg) for 7 days. For transplantation, 3×10⁷ brown preadipocytes were subcutaneously injected at the base of sternum on two sides in 8-week old male nude mice. Fat pads were carefully dissected after 2 weeks for gene expression analyses.