Murine mature B Cells: Control vs. Notch1 Signaling Activated. Murine mature B Cells: Control vs. Notch1 Signaling Activated

Transcriptional profiling of murine splenic mature B cells that have experienced activation-induced cytidine deaminase (Aicda) gene expression comparing control untreated B cells with B cells in which transgenic Notch1 intracellular domain (NICD1) gene, coding for an active form of Notch1, began to be expressed after Aicda promoter-driven expression of Cre recombinase. B cells that have experienced Aicda promoter activation were visualized by Cre-inducible expression of red fluorescent protein (RFP) which was inserted in the Rosa26 locus. Total RNA was extracted from splenic CD19+ RFP+ B cells isolated from mice using an RNeasy Mini kit (Qiagen, Hilden, Germany). Equal amounts of total RNA derived from 3 control mice and 3 mice with conditional activation of Notch1 signaling in mature B cells were pooled and submitted for gene expression microarray analysis. Goal was to determine the effects of conditional activation of Notch1 signaling in mature B cells. Overall design: Two-condition experiment, mature B cells vs. Notch1-activated mature B cells. Biological replicates: Equal amounts of total RNA from 3 mice per group were pooled together.

本数据集针对曾发生活化诱导胞苷脱氨酶（Aicda）基因表达的小鼠脾脏成熟B细胞开展转录谱分析，对比未处理的对照B细胞与经Aicda启动子驱动的Cre重组酶表达后，开始表达编码活化型Notch1的转基因Notch1胞内结构域（NICD1）基因的B细胞。通过插入于Rosa26位点的Cre诱导型红色荧光蛋白（RFP）的表达，对曾发生Aicda启动子激活的B细胞进行可视化标记。采用RNeasy Mini试剂盒（德国希尔德市Qiagen公司），从小鼠脾脏分离的CD19阳性、RFP阳性B细胞中提取总RNA。将3只对照组小鼠与3只成熟B细胞中存在Notch1信号条件性激活的小鼠的总RNA按等量混合后，提交进行基因表达微阵列分析。本实验旨在探究成熟B细胞中Notch1信号条件性激活的生物学效应。实验设计：双条件对照实验，即成熟B细胞组与Notch1激活的成熟B细胞组。生物学重复：每组各取3只小鼠的总RNA按等量混合后进行检测。