B6 cg lepob j

All animal experiments were performed according to procedures approved by Beth Israel Deaconess Medical Center’s Institutional Animal Care and Use Committee. The mice (strain: B6.Cg-Lepob/J from Jackson Labs) were 8 weeks old when administered inhibitor or vehicle by once daily oral gavage for 5 days. Doses in mice (GSK1120212, 3 mg/kg in DMSO; PD0325901, 10 mg/kg in DMSO) were diluted in 15% Cremophor EL/ 82% saline/ 3% drug or DMSO were comparable to that given in human clinical trials [5 (link), 9 (link), 10 (link), 14 (link)]. Kidneys, livers, and pancreata from nine mice were dissected and flash frozen in liquid nitrogen.

All animal care and experimental procedures were performed under approval of animal care committees of Harvard University. Male mice with genetic obesity were obtained from Jackson Laboratories (Strain: B6.Cg-Lep^ob/J, stock number: 000632) at 9–12 weeks of age and kept on regular chow diet. Male mice for the diet induced obesity model (HFD) were also obtained from Jackson Laboratories (Strain: C57BL/6J DIO, stock number: 380050), which were fed ad libitum for 20 weeks with 60% kcal fat diet (D12492i, Research Diets) after weaning. The mice were kept on a 12 hour day/night cycle and azoramide compound was fed to mice once a day at 3:00 pm by oral gavage (150mg/kg in 200 µl vehicle solution). The recipe for the vehicle solution was 10% (vol/vol) ethanol, 0.1%(vol/vol) Acetic Acid, 40% (vol/vol) PEG400 and 0.05% (weight/vol) CMC (carboxymethylcellulose). Glucose and weight measurements were taken at 3:00pm, after 6 hours food withdrawal. Glucose tolerance tests (GTT) were performed by intraperitoneal glucose injection (1 g/kg) following an overnight food withdrawal as described previously (18 (link)). Metabolic cage and euglycemic hyperinsulinemic clamp studies were performed as described previously (70 (link), 71 (link)).

All animal experiments were performed according to procedures approved by Beth Israel Deaconess Medical Center’s Institutional Animal Care and Use Committee, animal protocol number: 104-2014. The mice (strain: B6.Cg-Lepob/J from Jackson Labs) were 8 weeks old when administered nicotine in drinking water (200μg/mL) for 21 days. Brain, heart, kidney, liver, lung, pancreas, and spleen from 10 mice (5 nicotine-treated and 5 controls) were dissected and flash frozen in liquid nitrogen.

Wild type C57BL/6 mice were purchased from Harlan Sprague Dawley. Leptin deficient mice (ob/ob) were purchased from the Jackson Laboratory (B6.Cg-Lep^ob/J, stock No: 000632). These mice are diabetic, obese, and exhibit delayed wound closure [35 (link)-38 (link)]. Mice were housed in a centralized animal facility at the University of Chicago. Procedures were approved by the University of Chicago Institutional Animal Care and Use Committee, and the NIH guidelines for the care and use of laboratory animals (NIH Publication #85-23 Rev. 1985) were observed. Immunizations in C57BL/6 mice were performed subcutaneously with one 100 μL injection of 2 mM ovaQ11 in PBS followed by boosting at 4 weeks and 6 weeks with half doses (50 μL of 2 mM ovaQ11). Owing to difficulties in placing subcutaneous injections in the obese ob/ob mice, for this strain immunizations were performed via intraperitoneal injections with one 200 μL injection of either ovaQ11 or CFA-ovaQ11 and boosted with half doses of 100 μL of ovaQ11 or IFA-ovaQ11 respectively at 4 weeks and 6 weeks. Mice in “PBS” negative control groups received PBS injections of the same volume as the experimental groups. Serum samples were collected via the submandibular vein and analyzed by ELISA.

All studies were performed at the VA Palo Alto Health Care System, Palo Alto, CA. All animal experiments were performed per the procedures approved by the VA Palo Alto Health Care System Institutional Animal Care and Use Committee.
Seven to eight-week-old apoE KO mice (on C57BL/6J background strain B6.129P2-Apoetm1Unc/J stock number 002052) were purchased from Jackson Laboratory, Bar Harbor, ME. After arrival, these mice were acclimated for one week on pelleted laboratory rodent chow (Harlan, 2018) and later switched to a high-fat/Western diet (TD.88137, Harlan Teklad, Indianapolis, IN) ad libitum with continuously available water and maintained on this diet for up to 24 weeks. Seven to eight weeks old C57BL/6J (Stock No: 000664 | Black 6) were also obtained from Jackson Laboratory and either used directly in experimentation or used in the DIO model. To produce DIO mice, WT mice were fed a high-fat diet (#TD 06414, Envigo RMS, Indianapolis, IN) starting at 7–8 weeks of age and maintained on this diet for up to 24–26 weeks of age. ob/ob (B6.Cg-Lep^ob/J; Stock No: 000632 |B6 ob) were supplied by Jackson Laboratory at 7–8 weeks of age and used starting at 10 weeks of age. All animals were treated with vehicle or specific peptides as specified under each figure.