Lepob ob

All animal work was approved by the Yale University Institutional Animal Care and Use Committee. All mice used in this report were female. Mice were housed at 22–24°C under a 12 h light–dark cycle and were fed with regular chow (RC) (Harlan Teklad no. 2018, IN, USA; 18% energy from fat) or high-fat diet (HFD) (Research Diets, NJ, USA, D12451; 45% energy from fat); water was provided ad libitum. C57BL/6J (Jax, CT, USA, 000664), HAS-Cre79 (Jax, 006149) and Lep^ob/ob (Jax, 000632) mice were purchased from the Jackson Laboratory. The Tet3^fl/fl mice were generous gifts from A. Rao from La Jolla Institute for Immunology (CA, USA). The muscle-specific Tet3 knockout (mKD) mice were created by crossing Tet3^fl/fl mice with mice expressing Cre recombinase under the control of human α skeletal muscle actin (HSA) promoter (HSA-Cre79 mice). TET3 floxed littermates (wild-type [WT]) were used as controls. Randomisation was not feasible during group assignment, but results were analysed in a blinded manner whenever possible. Data were not included if values were excluded by outlier test. For information on animal numbers, refer to figure legends. Before experiments, mice were allowed to acclimate for at least 7 days in our animal facility.

Ten-month-old females Lep^ob/ob (The Jackson Laboratory; stock #000632) and 12-month-old females LepR^Null/Null mice (The Jackson Laboratory; stock #018989) were produced in the University of São Paulo. Lep^ob/ob mice were generated by crossing heterozygous B6.Cg-Lep^ob/J mice. The brains of males and females are substantially different^{41 (link),42 (link)} and cannot be directly compared. Therefore, considering the poor availability of information about female mouse brains in the literature, the current study focuses on female mice. Moreover, due to the high mortality of LepR^Null/Null mice after 9 months of age, the sample size for this experimental group is smaller than for the other groups in the current study.
Homozygotes were used as the spontaneous model of obesity, and 10- to 12-month-old WT mice (C57Bl/6J) littermate were used as WT controls. All mice were housed in climate-controlled rooms on a 12-h light–dark cycle, with standard rodent chow and water access ad libitum. All procedures were conducted in accordance with the Guiding Principles for the Care and Use of Research Animals. Animals were euthanized in compliance with the recommendations of the Animal Care Committee for the Federal University of Rio de Janeiro (Rio de Janeiro, Brazil) Process number: 01200.001568/2013-87.

All animal procedures were approved by Augusta University’s Institutional Animal Care and Use Committee (IACUC). Mice were maintained under standard conditions with controlled 12 h/12 h-light–dark cycle and 21 ± 1 °C room temperature. Lep^ob/ob, Lepr^db/db and C57BL/6 J mice were obtained from Jackson Laboratory (Bar Harbor, ME). Fam13a^−/− mice were backcrossed to C57BL/6 (Stock#: 027, Charles River) for seven generations and genotyped as previously reported [14 (link)]. 4–5 week old Fam13a^+/+ and Fam13a^−/− littermates were fed with high fat diet (HFD, D12492, 60% Kcal from fat, Research Diets, NJ) for up to 12 weeks. Mice were not randomized. Six week old C57BL/6 J male mice were simply randomized into two experimental groups for ad libitum access to low fat control diet (LFD, D12450B, 10% Kcal from fat, Research Diets, NJ) and 60% HFD for up to 12 weeks as reported [16 (link)]. All mice studies were not blinded. All mice were sacrificed after 4 h fasting with isoflurane.