D12492

In the previous study[7 (link)], body weight under chow diet (fat 4.5%, Lab Diets 5001) was recorded to 20 weeks of age, magnetic resonance imaging (MRI) was applied for body composition measurement at 20 weeks of age. For high fat diet feeding, four-week-old WT and TLR5KO mice were given a high-fat diet (fat 34.9%, D12492, Research Diets Inc., New Brunswick, NJ) for 8 weeks starting from 4 weeks of age. To avoid the confounding effects of co-housing on the diversity of cecal bacteria, mice from multiple litters that were housed separately were selected for gut microbiome analysis. In the current study, body weight under chow diet was recorded to 20 weeks of age, for high fat feeding, mice were given a high-fat diet (fat 34.9%, D12492, Research Diets Inc., New Brunswick, NJ) for 12 weeks starting from 8 weeks of age. Quantitative magnetic resonance imaging (QMR) was applied for body composition measurement at 20 weeks of age. To avoid the effects of co-housing on gut microbiome, multiple litters that were separately caged since weaning were selected for gut microbiome analysis.

All animal experimentation was approved and conducted in accordance with the guidelines from Hong Kong Baptist University and was endorsed by the University Human and Animal Subject Committee and the Department of Health, the Government of Hong Kong Special Administration Region. Male nude mice of 4 weeks old were purchased from the Chinese University of Hong Kong. DU145 cells (ATCC) were washed in phosphate-buffered saline (PBS) containing 5 mM EDTA, the cell number and viability were examined using trypan blue. Single-cell suspension of >90% viability was suspended at 1 × 10⁵ cells in 100 µl PBS and inoculated subcutaneously into each mouse. When the tumor grows to ~80 mm³ in size, mice were randomly selected to have either HFD (D12492, 60 kcal% fat, Research Diets) or the corresponding matched CD (D12450J, 10 kcal% fat 7% sucrose, a matched control for D12492, Research Diets). Alternatively, they had a diet rich in PA (D16042106) or the corresponding matched CD (D17041705). All these diets are formulated by Research Diets, Inc. The fatty acid compositions of these diets are shown in Supplementary Table S1. STA treatment^{22 (link)} was given at e dose of 2.5 mg/kg when the dietary intervention started. Both diet and water were supplied ad libitum. Body weight and tumor size of the xenograft mouse models were measured every day.

C57BL6 male mice were purchased from The Jackson Laboratory at 4-week of age and acclimated for 2-week. Mice were housed in a temperature-controlled environment with a 12 h light–dark cycle and ad libitum water and diet. At 6-week of age mice were fed either low fat diet (Research diet, D12450B) or high fat diet (Research Diets, D12492) for 13 weeks. During the last 6 weeks of the diet, the mice were intraperitoneally (i.p.) injected daily with roscovitine (50 mg/kg) or vehicle. For diet studies, 6-week-old C57BL/6 N wild-type male mice were fed a diet with 60% kcal% fat (high fat diet, Research Diets, D12492) for 9 weeks. All animal studies were approved by the Boston University School of Medicine Institutional Animal Care and Use Committee.