Adiponectin creer mice

To create MED19-AKO mice, we obtained mice with conditional-potential allele of MED19 (MED19^{tm1a(EUCOMM)Wtsi} from the EUCOMM repository and crossed them with actin-Flp transgenic mice to remove the targeting cassette and then subsequently crossed the mice with adiponectin-Cre mice obtained from the Jackson Laboratory (028020) to generate adipose-specific MED19 knockout (MED19-AKO) mice. To generate adipose-specific, tamoxifen-inducible MED19 KO (MED19-iAKO) mice, MED19 floxed animals were crossed with Adiponectin-CreER mice obtained from the Jackson Laboratory (025124). To induce knockout, 12-week-od control (MED19^Lox/Lox without Cre) and MED19-iAKO mice were treated daily with tamoxifen (100 μg/g body weight) for five consecutive days. All experiments used male mice unless otherwise stated. All mice were kept on a 12h light/dark cycle and provided ad libitum access to food (Purina 5053) and water. All animal experiments were performed in accordance with procedures approved by the Institutional Animal Care and Use Committee at Washington University School of Medicine.

All experiments conducted on mice were done following the guidelines issued by Yale’s University Institutional Animal Care and Use Committee (IACUC). C57/Bl6 mice were purchased from Charles River Laboratories and used at least one week after their arrival. Pdgfrα-Cre (C57BL/6-tg(PdgfRa-cre01Clc/J, stock #013148), mT/mG (B6.129(Cg)–Gt(ROSA)26Sortm4(ACTB–tdTomato,– EGFP)Luo/J, stock # 007676; Pdgfrα^fl (B6.Cg-Pdgfratm8Sor/EiJ, stock # 006492); Rosa-CreER (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, stock # 008463) were purchased from Jackson Laboratories. Adiponectin-CreER mice were provided by E. Rosen (Beth Israel Deaconess Medical Center, Boston, MA, USA) and are now available at Jackson Laboratories (stock # 024671). Pdgfa^fl mice were a generous gift from Christer Betsholtz (Uppsala University, Sweden). Aged mice (18, 24 and 30 month old) were obtained from the NIA. Akt2 KO mice were a generous gift from W. Sessa (Yale University, New Haven, CT, USA). A 30mg/ml tamoxifen stock (Sigma T5648) was generated in 100μl of ethanol (200 proof; Decon Laboratories #2716) and 900μl sesame oil (Sigma S3547) and administered at 100mg/kg/day via intraperitoneal injection. For in vivo proliferation assays mice were injected with 5-ethynyl-2′-deoxyuridine (EdU) (Invitrogen A10044) by intraperitoneal injection using a 50mg/kg of weight dose.