Goto kakizaki rats

All animal experiments were carried out in accordance with the Guide for the Care and Use of Laboratory Animals prepared by the National Academy of Sciences and published by the NIH, and were approved by the University of Cincinnati Animal Care and Use Committee (Animal Welfare Assurance Number: A3295-01). Goto-Kakizaki (GK) rats (10–11 weeks of age, male) with higher fasting and post-prandial blood glucose levels and age-matched normal male Wistar rats were purchased from Charles River Laboratories (www.criver.com). These rats were used for isolation and culture of primary cardiomyocytes, as described previously [22 (link)]. Briefly, cardiomyocytes were seeded into dishes pre-coated with mouse laminin (10 μg/ml) for 2 h at room temperature, followed by infection with Ad.rno-miR-320-off (#mr5309, Abm Inc.) for 48 h to knock down endogenous miR-320. Ad.miR-off (#m010, Abm Inc.) was used as a control. Culture supernatants were then collected for isolation of exosomes. A mouse cardiac endothelial cell line (MCEC) was initially developed in Dr. Weksler’s lab [23 (link)]. We purchased it from CELLutions Biosystems Inc. MCECs were cultured in DMEM with 10 mM penicillin/streptomycin (Gibco), 10 mM HEPES (Sigma) and 5% FBS in 0.2% gelatin-coated plates. For the exosome collection and function assays, exosome-depleted FBS (Syetem Biosciences Inc.) was used in the cell culture.