Gene expression in the islets of 10 day old, 4 week old and 12 week old Non-Obese Diabetic (NOD) mice vs. healthy, age-matched NOD.B10 control mice

Gene expression in the islets of NOD vs. NOD.B10 mice at 10 days, 4 weeks and 12 weeks of age insulitis. Pancreatic islets from 10 day old, 4 week old and 12 week old NOD and NOD.B10 mice were isolated by collagenase digestion and dissolved in 0.5 ml Trizol reagent (Invitrogen). 0.1 ml of chloroform was added. Samples were shaken vigorously and centrifuged at 12,000xg at 4°C. The clear aqueous phase was combined with an equal volume of 70% ethanol, and bound to a RNeasy micro column (Qiagen). RNA was extracted according to manufacturer’s instructions and RNA quality was assessed using the Agilent 2100 Bioanalyzer and the RNA 6000 Pico Reagent Kit (Agilent). Total RNA (300 ng) extracted from the islets of NOD mice were labelled with Cy5 and run individually against a pool of Cy3-labeled total RNA (300 ng) isolated from the islets of age-matched NOD.B10 mice (pool of 6 animals). Microarrays were performed using the Whole Mouse Genome Microarray Kit, 4×44K 2-color arrays (Agilent Technologies).