C57bl 6j dio mice

All mouse care and experimental procedures were approved by the institutional animal care research advisory committee of the Albert Einstein College of Medicine. All experiments were performed in accordance with relevant guidelines and regulations. Mice used in experiments included C57BL/6J mice (The Jackson Laboratory, stock # 000664) and C57BL/6J DIO mice used as noted (the Jackson Laboratory, Stock # 380050). Both male and female mice were used for most experiments and were maintained with a 12 hours light-dark cycle. C57BL/6J mice at 5 weeks of age were fed a low fat diet (LFD, 70% calories provided by carbohydrates, 20% by protein, and 10% by fat; 3.85 kcal/g, D12492J, Research Diets), or a high fat diet (HFD, 20% calories by carbohydrate, 20% by protein, and 60% by fat; 5.21 kcal/g, D12492; Research Diets) with ad libitum access to water for 5 weeks at 28°C. Mice were euthanized by an overdose of Isoflurane.

All animal care protocols have been approved by UAMS Institutional Animal Care and Use Committee AUP #3788. C57BL/6J DIO mice obtained from Jackson Laboratory (Bar Harbor, ME) were housed in polycarbonate cages on a 12-h light-dark cycle at University of Arkansas for Medical Sciences. Diets were purchased from Research Diets, Inc. and administered to mice starting at 8 weeks of age. Mice were administered a palm oil-based HF diet D15012001 (4.70 kcal/g, 45 kcal% fat^{13 (link)}) and control diet D12450B (3.85 kcal/g, 10 kcal% fat) for 14 weeks. In a separate study 8-week-old mice were treated as follows: (a) one group of mice received the control diet (D12450B); (b) two groups of mice were administered the palm oil-based HF diet D15012001. Two weeks prior to harvesting tissues (week 11.5–14), the drinking water of one group of mice kept on the 45% HF diet was replaced with a solution containing 30 mM PBA. The other two groups of mice treated with control diet and palm oil-based HF diet had 30 mM NaCl in their drinking water.

32, 21 week old, male C57BL6/J DIO mice were obtained from the Jackson Laboratories (Bar Harbor, ME), fed 60% high fat diet (Research Diets D12492). Mice were acclimatized for 2 weeks at 72 °F (ambient) then moved to 80 °F that is close to thermoneutral zone, to acclimate another week prior to baseline randomization and subsequent surgery. For experiments at ambient temperature, mice were housed at 72 °F throughout the study. Animals were allocated into 4 groups of n = 8 based on BW and glucose levels to achieve equal distribution of these parameters in each group. On day 0, all animals underwent surgery for the implantation of an Alzet® mini-pump (Model #2002, Durect Corporation) for continuous delivery of either vehicle (PBS) or 0.85 mg/kg/day native FGF21. The iBAT was surgically removed at the time of mini-pump implant in those mice allocated into the X-BAT groups. VO₂ and VCO₂ were measured for 24 hours of fed and 24 hours of fasted conditions using a comprehensive laboratory animal monitoring system (CLAMS) equipped with an Oxymax Open Circuit Calorimeter (Columbus Instruments, Columbus, OH) as previously described (n = 8/group)20 (link)30 (link). RT-PCR One-Step (Qiagen) was used to determine the expression of UCP-1 by standard PCR protocol.

Male C57BL/6J DIO mice (Jackson Lab # 380050, Bar Harbor, ME, USA) and their respective controls (Jackson Lab # 380056) received either a rodent diet 60 kcal% fat (Research Diets D12492, Research Diets Inc., New Brunswick, NJ, USA) or a 10 kcal% fat (Research Diets D12450B) from 6 weeks of age to induce obesity. The mice had free access to their respective diet and water, were kept in a room with 12 h/12 h dark/light cycle, and were sacrificed at 15–19 weeks. Experiments were conducted based on the guidelines of the National Institutes of Health Guide for the Care and Use of Laboratory Animals and approved by Tulane University’s Institutional Animal Care and Use Committee (Protocol 1398).

Male C57BL/6 J DIO mice fed with a high-fat diet (HFD, 60% of kcal from fat) since 6 weeks of age were purchased from Jackson Laboratories (Bar Harbor, ME). DIO mice were acclimated at Rigel for 4 weeks prior to study initiation. Mice were housed 4 per cage in a room with a 12-12 hour light/dark cycle and were given HFD (D12492, Research Diets, Inc., New Brunswick, NJ) and drinking water ad libitum. Rigel’s Institutional Animal Care and Use Committee approved all procedures (IACUC Protocol No. Rigel 2-2012).

All experiments performed in mice were approved by the Institutional Animal Care and Use Committee at the Washington University School of Medicine. Liver-specific Mpc2 knockout (LS-Mpc2−/−) or cardiac-specific Mpc2 knockout (CS-Mpc2−/−) mice were generated as described using LoxP technology with Cre expression driven by the albumin gene promoter (16 (link)) or myosin light chain 2v promoter (25 (link)), respectively. Control mice were littermates not expressing Cre recombinase (fl/fl).
For HF diet studies, LS-Mpc2−/− and littermate control mice were switched from standard chow to a 60% HF diet (Research Diets Inc, #D12492) at 7 to 8 weeks old, and experiments were performed after 12 weeks on diet. For studies not in the Mpc2 null background, C57BL/6J DIO mice were purchased from Jackson Laboratory (cat #380050) after 9 weeks of feeding a 60% fat diet (Research Diets Inc, #D12492) and were maintained on the same diet for the indicated times. When included in the experimental design, low-fat (LF) diet C57BL/6J control mice were also purchased from Jackson Laboratory (cat #380056) and fed 10% LF control diet (Research Diets Inc, #D12450B). LF diet controls were matched in age to DIO comparators.