Gene expression by cyotosolic DNA stimulation. Mus musculus

STING molecule has been reported to be important adaptor molecule for cytosolic DNA sensing. We investigated gene expression by cytosolic DNA stimulation using bone marrow derived dendritic cells. We comparared gene expression profile between WT and STING knock out BMDCs after cytosolic DNA stimulation. Overall design: We generated BMDCs by culturing in the presence of GM-CSF. BMDCs were stimulated by purified tumor-cell derived DNA using lipofectamine. After 7 hrs incubation, total RNA was isolated using Rneasy kit (Qiagen) and microarray was performed at genomic core facility in university of Chicago

STING分子（STING）已被报道为胞质DNA感知通路的关键接头分子。本研究以骨髓来源树突状细胞（BMDCs）为实验模型，通过胞质DNA刺激开展基因表达检测。我们对比了野生型（WT）与STING基因敲除骨髓来源树突状细胞在胞质DNA刺激后的基因表达谱。
实验设计概述：本研究通过在含有粒细胞-巨噬细胞集落刺激因子（GM-CSF）的培养基中培养诱导生成骨髓来源树突状细胞。采用Lipofectamine脂质转染试剂，将纯化的肿瘤细胞来源DNA转染刺激骨髓来源树突状细胞。孵育7小时后，使用RNeasy试剂盒（Qiagen）提取总RNA，并于芝加哥大学基因组核心设施完成微阵列芯片检测。