High-fat, high-carb diet in mice

C57Bl/6 mice pups (37 of each sex) were obtained from Harlan Laboratories S.R.L. (S.Piero al Natisone, Italy). Immediately after weaning, animals were randomly group-housed in cages in a temperature-controlled environment (22±2°C) and on a 12 hours light/dark schedule, and fed ad-libitum with control diet (CTRL, D12328, Research Diets, New Brunswick, NJ) or HFHC diet (HFHC: D12331, Research Diets, New Brunswick, NJ), plus 42g/L fructose/sucrose in drinking water, for a total of 16 weeks [25 (link)]. Four experimental checkpoints were established (4, 8, 12 and 16 weeks of diet), in which part of the animals was sacrificed. Animal care and procedures were conducted in accordance with the Italian Law (decree 116–92) and by European Community directive 86-609-EEC. All experimental protocol were approved by the local committee of the Medical Research Institute and by the National Authority (Ministero della Salute—Direzione generale della sanità animale e dei farmaci veterinari—Ufficio VI, Rome, Italy, under the Article 7, D.lgs 116/92). The maximal effort was done to reduce the number of animals used and their sufferance in the respect of the 3R rule.