2.1. Animal preparation and DCM model
This protocol is extracted from research article:
Kir6.1 improves cardiac dysfunction in diabetic cardiomyopathy via the AKT‐FoxO1 signalling pathway
J Cell Mol Med, Feb 6, 2021; DOI: 10.1111/jcmm.16346

Male pathogen‐free C57BL/6J mice at 5 weeks of age were supplied by the Experimental Animal Center of PLA General Hospital. The study protocols were approved by the Ethics Committee of Chinese PLA General Hospital. All the mice were treated in strict accordance with the NIH guidelines or the guidelines from Directive 2010/63/EU of the European Parliament on the protection of animals used for scientific purposes. DCM was induced using a HFD (D12492; Research Diets, New Brunswick, NJ, USA) together with an single intraperitoneal injection of streptozotocin (STZ; Sigma‐Aldrich, St. Louis, MO, USA). 14 The mice were randomly divided into two groups: the DCM group was fed an HFD for 4 weeks, injected with STZ (100 μg/g of bodyweight) and then fed with HFD for another 12 weeks; the control group was fed a regular diet (D12450J, Research Diets, New Brunswick, NJ, USA) and injected with the same volume of vehicle (0.1 mol/L sodium citrate, Sigma‐Aldrich). Mice were housed five per cage, with free access to food and water. The HFD (5.21 kcal/g) consists of 60% calories from fat, 20% carbohydrate and 20% protein and the regular diet (3.82 kcal/g) contains 10% fat, 70% carbohydrate and 20% protein. The data regarding food intake and caloric intake for each experimental group are listed in Table S1. Plasma glucose levels were measured at the beginning and 12 weeks after the STZ injection by a Contour glucose meter (Roche, Basel, Switzerland). Mice with a fasting plasma glucose of over 13.89 mmol/L were considered diabetic (Figures S1 and S2). Mouse serum samples were analysed for Insulin using commercial enzyme‐linked immunosorbent assay (ELISA) kits (Ray Biotech, Norcross, GA, USA) according to the manufacturer's instructions. The data of insulin sensitivity test for all animals are provided in Figure S3. The mice were killed with an anaesthetic overdose of pentobarbital (100 mg/kg of bodyweight, Sigma‐Aldrich) injected intraperitoneally to obtain their samples.

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