JmjD2d-dependence of LPS-induced genes. Mus musculus

In dendritic cells, expression of the H3K9me3 demethylase JmjD2d is upregulated by LPS stimulation. To identify genes whose induction by LPS depends on JmjD2d activity, we performed a microarray analysis of wild-type and JmjD2d-knockdown dendritic cells, before and after stimulation with LPS. Overall design: RNA was extracted from three independent cultures of wild-type and JmjD2d-knockdown dendritic cells, before and 4h after stimulation with 100ng/ml LPS. The unstimulated and stimulated wild-type samples, and the stimulated JmjD2d-knockdown samples, were used for microarray analysis.在树突状细胞（dendritic cells）中，脂多糖（lipopolysaccharide，LPS）刺激可上调H3K9me3去甲基化酶JmjD2d的表达。为鉴定受LPS诱导且依赖JmjD2d活性的基因，我们对野生型与JmjD2d敲低的树突状细胞在LPS刺激前后进行了微阵列芯片（microarray）分析。整体实验设计如下：我们从3批独立培养的野生型及JmjD2d敲低树突状细胞中提取RNA，分别采集未受刺激以及经100ng/ml LPS刺激4小时后的样本；未刺激野生型样本、刺激后野生型样本与刺激后JmjD2d敲低样本均被用于本次微阵列芯片分析。