Genome wide expression analysis of high passage growth arrested BJ fibroblasts maintained or withdrawn from rapamycin maintenance.

BJ fibroblasts were grown in the presence of rapamycin until reaching growth arrest at high passages, at which point cells were maintained in or withdrawn from rapamycin. RNA was then harvested for RNA-seq Overall design: BJ fibroblasts were grown to growth arrest in the presence of rapamycin maintenance. Cells were then either maintained in rapamycin or withdrawn from rapamycin for two weeks. Experiments, excepting control treatment, were done in duplicates. RNA-seq tag density was calculated over gene bodies, and each withdrawn samples was compared to its paired maintained sample to determine fold change in gene epression.

将BJ成纤维细胞（BJ fibroblasts）置于添加雷帕霉素（rapamycin）的培养基中培养，直至其在高代次时进入生长阻滞状态；随后将细胞分为两组，一组继续在含雷帕霉素的环境中维持培养，另一组则撤除雷帕霉素。之后收集细胞RNA用于RNA测序（RNA-seq）。实验整体设计：将BJ成纤维细胞置于添加雷帕霉素的培养基中培养至生长阻滞状态，随后将细胞分别维持于雷帕霉素环境中，或撤除雷帕霉素并持续培养两周。除对照处理外，其余实验均设置双重复样本。通过计算基因区域内的RNA-seq标签密度，将每一组撤除雷帕霉素的样本与其配对的维持培养样本进行比较，以确定基因表达的倍数变化。