Gene expression in the pancreatic lymph nodes (PLN) of hyperglycemic 16-week-old Non-Obese Diabetic (NOD) mice compared to expression the PLN of euglycemic 16-week-old NOD mice

These experiments were performed to identify differentially expressed genes in the pancreatic lymph nodes of hyperglycemic NOD mice with high levels of destructive insulitis compared to euglycemic mice with lower levels of insulitis. The pancreatic lymph nodes of 16-week-old hyperglycemic and euglycemic NOD mice were isolated and homogenized in Trizol reagent (Invitrogen). Total RNA was extracted from the aqueous phase using the RNeasy mini kit (Qiagen). RNA quality was assessed using the Agilent 2100 Bioanalyzer and the RNA 6000 Nano Reagent Kit (Agilent). Total RNA from the PLN of hyperglycemic mice (n=2) were labelled with Cy5 and run individually against a pool of Cy3-labeled total RNA isolated from euglycemic mice (n=2). Microarrays were performed using the Whole Mouse Genome Microarray Kit, 4x44K 2-color arrays (Agilent Technologies).本实验旨在鉴定高血糖且伴重度破坏性胰岛炎的非肥胖糖尿病（NOD）小鼠与低糖岛炎水平的正常血糖NOD小鼠的胰腺淋巴结中差异表达基因。分离16周龄高血糖与正常血糖NOD小鼠的胰腺淋巴结，于Trizol试剂（Invitrogen）中匀浆；采用RNeasy小型试剂盒（Qiagen）从水相中提取总RNA；使用安捷伦2100生物分析仪及RNA 6000 Nano试剂盒（Agilent）评估RNA质量；将2只高血糖小鼠胰腺淋巴结的总RNA（n=2）以Cy5标记，分别与2只正常血糖小鼠分离得到的Cy3标记总RNA混合池进行杂交实验；采用安捷伦科技的全小鼠基因组4x44K双色微阵列试剂盒完成微阵列实验。