C57bl 6j mice

Animal husbandry and experiments with mice were performed in accordance with the Regulation of Animal Experiments of the National Center for Global Health and Medicine (Tokyo, Japan), and were approved by the Institutional Animal Care and Use Committee of the National Center for Global Health and Medicine. Male C57BL/6J mice (CLEA Japan, Tokyo, Japan) and db/db (C57BLKS/J Iar– + Lepr^db/+Lepr^db) and db/m (C57BLKS/J Iar– + m^db/+Lepr^db) mice obtained from the Institute for Animal Reproduction were used for experiments at 8 weeks of age. For the high-fat diet, C57BL/6J mice were fed chow containing 30% fat by weight (14% bovine fat, 14% porcine fat, and 2% soybean oil; Oriental Yeast Co., Tokyo, Japan) from 4 to 24 weeks of age. Mice heterozygous for a floxed Phd3 allele were generated by injecting Phd3 floxed embryonic stem cells into C57BL/6N blastocysts. Phd3-targeted embryonic stem cell clones were obtained from Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (Augsburg, Germany). To generate liver-specific Phd3 knockout (KO) mice, we intercrossed mice homozygous for a floxed Phd3 allele (F/F mice) with α1 anti-trypsin-Cre mice^{3 (link)}. Male F/F and KO mice were fed normal chow. Primary hepatocytes were prepared from these mice at 8 to 12 weeks of age.

All animal experimental procedures were reviewed and approved by the Institutional Animal Committee at the Department of Veterinary Science of Osaka University School of Medicine and performed in accordance with guidelines for animal experimentation at research institutes (Ministry of Education, Culture, Sports, Science and Technology, Japan), guidelines for animal experimentation at institutes (Ministry of Health, Labor and Welfare, Japan), and guidelines for the proper conduction of animal experiments (Science Council of Japan). Seven or eight-week-old male C57BL/6J mice and 8-week-old female C57BL/6N mice were purchased from CLEA Japan Inc. and housed in a temperature-, humidity- and light cycle-controlled facility (23 ± 1 °C; 55 ± 10%; light, 8:00–20:00; dark, 20:00–8:00). Mice had free access to food and water except for mice under pair-feeding condition. C57BL/6J mice were fed either a ND (MF, 12.8 kcal% fat; Oriental Yeast Co., Ltd) or a HFD (D12492, 60 kcal% fat; Research Diets Inc.), and C57BL/6N mice were fed a ND.

Animal care and all experiments were conducted in accordance with the guideline of the Animal Committee of Tokyo Medical and Dental University. C57BL/6J mice were purchased from ORIENTAL YEAST Co., Ltd. (Tokyo, Japan). Mice were housed under a 12-hour light-dark cycle and allowed food and water ad libitum. λ-Carrageenan was purchased from Sigma-Aldrich (St. Louis, MO). Recombinant mouse follistatin was purchased from R&D Systems Inc. (Minneapolis, MN). Rat anti-mouse monoclonal F4/80 antibody was purchased from AbD Serotec (Oxford, UK).

Homozygous (ADAR2^flox/flox/VAChT-Cre.Fast; AR2) and heterozygous (ADAR2^flox/+/VAChT-Cre.Fast; AR2H) conditional ADAR2 knockout mice were used in this study19 (link)20 (link). In these mice, Cre is selectively expressed in motor neurons under the control of the vesicular acetylcholine transporter promoter34 (link), ablating the ADAR2^flox gene in approximately 50% of motor neurons by the age of five postnatal weeks. As a result, 100% and no more than 30% of GluA2 is unedited at the Q/R site in the motor neurons of AR2 and AR2H mice, respectively. In these mice, the expression of GluA2 that is unedited at the Q/R site results in the slowly progressive death of motor neurons via Ca²⁺-permeable AMPA receptor-mediated mechanisms19 (link)20 (link). Both genders of AR2 and AR2H mice were used in this study. C57BL/6J mice (Oriental Yeast Co., Ltd.) of the same age were used as the WT control of the same strain. The mice were housed at two to three per cage on a 12 h: 12 h light-dark cycle with free access to food and water. All of the studies were approved by the Committee on Animal Handling at the University of Tokyo and were performed in accordance with the guidelines for animal experiments of the Ministry of Education, Culture, Sports, Science and Technology, Japan.