Microarray mRNA expression profiling in Stat5b knockdown pancreatic beta cell line. Mus musculus

Stat5b gene is located at IDD4 type 1 diabetes susceptibility interval, and is mutated in NOD mice. This mutation has been linked to diabetes with strong association with autoimmune aetiology. We have treated MIN6 pancreatic beta cell line with Stat5b siRNA and analyzed the differential expression profile in control siRNA treated versus Stat5b siRNA treated cells in an effort to understand the role of Stat5b in diabetes. Overall design: MIN6 cells were treated with control siRNA or Stat5b siRNA for 72 hours, followed by prolactin stimulation. The cells were trypsinized and total RNA isolated using miRNeasy Mini Kit (Qiagen). The mRNA differential expression was compared using Illumina microarray platform.

Stat5b基因定位于IDD4型1型糖尿病易感区间，并在非肥胖糖尿病（NOD）小鼠中发生突变。该突变与糖尿病存在强相关性，且与自身免疫病因学密切相关。为探究Stat5b在糖尿病中的作用，我们采用Stat5b小干扰RNA（siRNA）处理MIN6胰腺β细胞系，并对比对照组siRNA与Stat5b siRNA处理组细胞的差异表达谱。总体实验设计如下：MIN6细胞经对照siRNA或Stat5b siRNA处理72小时后，施加催乳素刺激；随后对细胞进行胰酶消化，使用miRNeasy Mini试剂盒（Qiagen）提取总RNA；采用Illumina微阵列平台完成mRNA差异表达分析。