Dlk1-Mediated Temporal regulation of Notch Signaling is Required for Differentiation of Alveolar Type II to Type I Cells during Lung Repair

Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived retinal transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis Methods: Lung epithelia cells were collected after 10 days of treatment by pseudomonas aeruginosa (PA), RNA-Seq and qRT-PCR were applied to analyze the high-throughput data Results: In general, the mutant type II-like cells expressed higher levels of type I cell markers and lower levels of type II cell markers Overall design: Total RNA profiles of mutant(SpC-CreER/Dlk1flox/ROSA-Tomato) and wild-type(SpC-CreER/ROSA-Tomato) mice were generated by deep sequencing, in triplicate, using Illumina HiSeq4000.

研究目的：下一代测序（Next-generation sequencing，NGS）已彻底革新了基于系统生物学的细胞通路分析。本研究旨在对比基于NGS的视网膜转录组分析（RNA-seq）与微阵列、定量反转录聚合酶链反应（qRT-PCR）的检测方法，并评估适用于最优高通量数据分析的实验方案。
实验方法：经铜绿假单胞菌（Pseudomonas aeruginosa，PA）处理10天后收集肺上皮细胞，采用RNA-seq与qRT-PCR对高通量数据进行分析。
实验结果：总体而言，突变型II型样细胞的I型细胞标志物表达水平更高，而II型细胞标志物的表达水平更低。
整体实验设计：本研究采用Illumina HiSeq4000平台对突变型（SpC-CreER/Dlk1flox/ROSA-Tomato）与野生型（SpC-CreER/ROSA-Tomato）小鼠的总RNA进行深度测序，每组设置3次生物学重复。