Lepr db db mice

All animal experiments were conducted at the Animal Facility of the AMC and approved by the Committee for Animal Welfare (DLV102141-1) of the AMC, Amsterdam, The Netherlands. Eleven-week old male Lepr^db/db mice and control Lepr^db/m mice from the same colony on the C57B/6J background were purchased from The Jackson Laboratory (Bar Harbor, USA). Animals were housed in a constant 12h light/dark cycle with controlled temperature and humidity and were given ad libitum access to water and food (Purina lab Diet #5008). At twelve weeks of age, the nonfasting animals were sacrificed and blood samples were obtained. Plasma was isolated by centrifugation for 15 minutes at 3,000 rpm at 4°C and plasma was stored at -80°C for future analysis. Plasma cholesterol and TG were measured with a colorimetric assay (Roche, Basel, Switzerland). Plasma glucose was analysed using strips from Contour (Bayer, Zurich, Switzerland) according to the manufacturers protocol and plasma insulin levels were measured using a commercially available ultrasensitive Elisa from Mercodia (Uppsala, Sweden). Tissues were snap-frozen into liquid nitrogen.

All animal experiments were performed following the approval and guidelines of the Institutional Animal Care and Use Committee of Indiana University. Animals are maintained in accordance with the applicable portions of the Animal Welfare act and the DHHS Guide for the Care and Use of Laboratory Animals. LIPTER(Tg) C57BL/6J mice were generated using CRISPR/Cas9 at the Genome Editing, Transgenic, and Virus Core at UPMC Magee-Women’s Research Institute. Six-week-old male homozygous Lepr^db/db mice were obtained from the Jackson Laboratory (strain number 000697). Myh10 floxed mice with C57BL/6J background were obtained from Dr Robert S. Adelstein lab at Laboratory of Molecular Cardiology, National Heart, Lung and Blood Institute^{57 (link)}. C57BL/6J Tnnt2^Cre mice were obtained from Dr Chenleng Cai lab at Indiana University. Conditional Myh10^cKO in mouse CMs was generated by crossing Myh10^f/f mice with Tnnt2^MerCreMer mice. Six-week-old male Myh10^f/f and Myh10^f/f/Tnnt2^MerCreMer mice were injected with tamoxifen (0.1 mg g⁻¹ body weight) for days 1, 3 and 5 by intraperitoneal injection and then fed with a HFD (45 kcal% fat, Research Diets Inc., D12451i) for 3 months. Six-week-old male LIPTER(Tg) mice and age-matched WT C57BL/6J mice were fed with a HFD (45 kcal% fat, Research Diets Inc., D12451i) or NC (18 protein, Inotiv, 2018sx) for 10 months.

All animal protocols were approved by the New York University School of Medicine Institutional Animal Care and Use Committee. We obtained male and female diabetic (Lepr^db/db) mice, aged 6–8 weeks, from The Jackson Laboratory (Bar Harbor, ME, USA). All Lepr^db/db mice used in the experiments had blood glucose concentrations of > 400 mg/dL. We obtained C57BL/6N wild type (WT) mice from Taconic Biosciences (Rensselaer, NY, USA). We housed these mice in a temperature-controlled, virus-free barrier animal facility with a 12-h light/12-h dark cycle and maintained them on chow diet and water ad libitum.