Teklad diet 2016

The animal experiments were approved by the Ethical Committee for Animal Research in Gothenburg (Göteborgs djurförsöksetiska nämnd; ethic numbers 136–16, 467–19, 4072–22). All experimental procedures and animal handling were performed and reported according to relevant guidelines and regulations, including ARRIVE guidelines. Homozygous C451A mice, with a point mutation at the C451 palmitoylation site in ERα^{11 (link),13 (link)}, and wildtype (WT) littermates were generated by breeding female ERαC451A^+/- mice with male ERαC451A^+/- mice. Primers 5ʼ-CTAAACAAGCTTCAGTGGCTCCTAG-3ʼ and 5ʼ-ACCTGCAGGGAGAAGAGTTTGTGGC-3ʼ were applied for genotyping. All animals were housed in a standard animal facility under controlled temperature (22 °C) and photoperiod (12 h light:12 h darkness cycle). Phytoestrogen-free pellet diet (Teklad diet 2016, Envigo) and tap water ad libitum were given to the mice.

All studies were carried out on adult male Ghsr-IRES-Cre heterozygous mice crossed (or not) with ROSA26-ZsGreen reporter mice B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J (The Jackson Laboratory; stock number 007906) as previously described (Mani et al., 2017 (link)). The original heterozygous colony was produced by Prof Jeffrey M. Zigman (UT Southwestern Medical Center, Dallas, United States) and the breeding pairs were kindly provided by Prof Zane B. Andrews (Monash University, Melbourne, Australia). The mice used in this study were bred in-house at Gothenburg University (Sweden).
Mice were housed in a 12-h light/dark cycle (lights on a 7:00) at 20 ± 2°C and 50% humidity and had ad libitum access to standard maintenance chow (Teklad diet 2016; Envigo, Cambridgeshire, United Kingdom) and water unless otherwise stated. All procedures were approved by the local Ethics Committee for Animal Care and Use at the University of Gothenburg (permit number: 27-2015 and 132-2016) and conducted in accordance with legal requirements of the European Community (Decree 86/609/EEC).