G albumin-cre transgenic mice to Ppard f/f mice. Ppara knockout mice (PPARKO), FVN/NJ and FVB/NJ-db/db mice were purchased from Jackson Laboratory. Animals had been on chow diet regime (using the exception of Extended Data Fig 4f,g) and housed in a barrier facility with 12hour light and dark cycles. ZT0: lights on at six am; ZT12: lights off at 6 pm. All animal studies were approved by the Harvard Medical Region Standing Committee on Animals. Adenovirus-mediated liver-specific over-expression of knockdown– Adenovirus was injected by way of the tail vein (109 pfu/mouse). Subsequent metabolic characterizations had been carried out four days post injection. AdPPAR/adGFP was repeated in three cohorts (eight?0 weeks old male, n=4?) and LACC1KD was carried out in 2 cohorts (8?0 weeks old male, n=5). Circadian gene expression–5 cohorts have been applied for circadian research (8 weeks old, four male and 1 female cohorts, showing comparable final results). For circadian studies, animals have been sacrificed every single four hours beginning at 10AM (ZT4) for 24 hours (n=3?/genotype/time point) with absolutely free access to food and water. For dark cycle time points, animals had been sacrificed beneath red security light prior to dissection. For daytime restricted feeding research, animals have been fed everyday involving 6AM (ZT0) and 2PM (ZT8) for 7 days under 12-hour light and dark cycles. On the 8th day, animals had been sacrificed every 4 hours beginning at 6AM (ZT0) for 24 hours (n=3/genotype/time point). GW501516 treatment–Wild-type and LPPARDKO mice (n=4?/genotype/treatment) have been gavage with 2mg/kg body weight/day GW501516 carried by 0.5 methylcellulose remedy for four days. Animals have been sacrificed four hours following the final gavage.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptPC(18:0/18:1) injection studies–For the pilot experiment, eight?0 week old male C57BL/6J mice were i.p. injected with 1.25? mg/kg Pc(18:0/18:1). Circulating lipids levels were determined two and four hr just after injection to figure out the biological activity and dosing for Computer(18:0/18:1). five mg/kg in 5 intralipid was later applied for tail vein injection in FVB/NJ and serum lipids have been measured 4 hr later. Pc(18:0/18:1) showed related lipid lowering effects when injection was performed in the course of the dark (ZT12) or light (ZT8) cycle. FVB/NJ mice were utilized for these research for technical causes (ease of tail vein injection).Nature. Author manuscript; obtainable in PMC 2014 August 22.1349151-98-9 uses Liu et al.PagePC(18:0/18:1) infusion studies–8?0 week old male C57BL/6J and PPARKO mice (n=6?/genotype/treatment) were catheterized by way of the jugular vein.Formula of Perfluoroundecanoic acid 5 days postoperation, animals had been infused with Pc(18:0/18:1) or car carried by 5 intralipid at a rate of 25 /kg/min for 200 minutes at ZT4 (10 am).PMID:26760947 After infusion, a bolus of 10 i 3Holeic acid was infused to identify the in vivo fatty acid uptake price as described inside the strategy section. db/db mice–Eight week old male FVB/NJ-db/db mice had been injected using a bolus of 5mg/kg body weight Computer(18:0/18:1) or car carried by 5 intralipid through tail vein once everyday for six days (n=4?/treatment), followed by metabolic research. Metabolic studies Metabolic cage research were performed within a Comprehensive Lab Animal Monitoring Technique (Columbus Instruments). Data had been collected for 48 hours beginning at the beginning of the dark cycle. TG and FFA have been determined by colorimetric solutions (Thermo and Wako). Hepatic TG was determined from chloroform:methanol (2:1 v/v) extracts of vacuum dried liver samples. Glucose (GTT) and insulin toleran.
