Lymph node stromal cells: Control siRNA treated vs. Eif4g3 siRNA treated. Mus musculus

Gene expression in the top, light and heavy polysome fractions of Eif4g3 siRNA treated lymph node stromal cells (LNSCs) compared to control-sIRNA treated samples This study was performed to examine whether the expression of certain genes in LNSCs is regulated by the translation factor, eukaryotic translation initiation factor 4 gamma 3 (Eif4g3). Overall design: LNSCs were grown in 10 cm plates, allowed to reach ≥ 80% confluency and then transfected with 400 pmol of control or Eif4g3 siRNA using Lipofectamine 2000. 7 plates were transfected with control or Eif4g3 siRNA. Cells were pooled, lysed and fractionated on a 10% to 60% continuous sucrose gradient. Fractions were collected and pooled into the top, light and heavy polysome fractions. RNA was extracted from these samples and used for microarrays on the Agilent Whole Mouse Genome Microarray Kit, 4×44K 2-color arrays.

本数据集为靶向真核翻译起始因子4γ3（Eukaryotic translation initiation factor 4 gamma 3, Eif4g3）的小干扰RNA（siRNA）处理的淋巴结基质细胞（Lymph Node Stromal Cells, LNSCs）的顶层、轻级及重级多聚核糖体组分中的基因表达，与对照siRNA处理样本的对比数据。本研究旨在探究LNSCs中部分基因的表达是否受翻译因子Eif4g3的调控。整体实验设计：将LNSCs接种于10 cm培养皿中，待细胞汇合度达到≥80%后，使用Lipofectamine 2000转染试剂将400 pmol的对照siRNA或Eif4g3 siRNA转染至细胞中。共7个培养皿分别转染对照siRNA或Eif4g3 siRNA。收集所有细胞并裂解，随后通过10%至60%的连续蔗糖密度梯度离心进行组分分离。收集各梯度组分并合并为顶层、轻级及重级多聚核糖体组分。从上述样本中提取RNA，采用安捷伦全小鼠基因组微阵列试剂盒（4×44K双色微阵列）进行基因芯片检测。