Prostaglandin E2 inhibits pro-fibrotic function of human pulmonary fibroblasts by disrupting Ca2+-signaling. Prostaglandin E2 inhibits pro-fibrotic function of human pulmonary fibroblasts by disrupting Ca2+-signaling

Purpose: Identification of gene expression in human lung fibroblasts, including analysis of impact of IPF (Idiopathic Pulmonary Fibrosis) as well as TGFβ stimulation, with particular emphasis on prostanoid receptors and related pathways. Methods: Truseq stranded mRNA sequencing, via Illumina Hiseq 4000. Single reads, 75bp, ~25 million reads per sample. Quantification of gene expression in TPM using Kallisto, followed by Tximport. mRNA isolated via Qiagen RNeasy kit, with on-column DNase1 digestion. Results: Quantification of gene expression in primary human lung fibroblasts, including quantification of prostanoid GPCRs. Overall design: Transcriptomes of human lung fibroblasts (HPFs), assayed via RNA-seq for HPFs with/wo TGFβ treatment, from normal donors and patients with IPF; n =3 for each group, with 4 groups.

### 研究目的
本数据集旨在对人类肺成纤维细胞的基因表达进行鉴定，包括分析特发性肺纤维化（Idiopathic Pulmonary Fibrosis, IPF）以及转化生长因子β（TGFβ）刺激的影响，重点聚焦于前列腺素受体及其相关通路。
### 实验方法
采用Illumina Hiseq 4000平台开展Truseq链特异性mRNA测序。测序模式为单端读长，读长75bp，每个样本约获取2500万条读段。使用Kallisto以每百万转录本（Transcripts Per Million, TPM）对基因表达量进行定量，随后通过Tximport进行后续数据处理。mRNA提取采用Qiagen RNeasy试剂盒，并辅以柱上脱氧核糖核酸酶I（DNase I）消化步骤。
### 实验结果
实现了原代人类肺成纤维细胞的基因表达定量分析，涵盖前列腺素类G蛋白偶联受体（GPCRs）的定量检测。
### 整体实验设计
通过RNA测序对人类肺成纤维细胞（HPFs）的转录组进行分析，受试样本分为4组：来自健康供体与特发性肺纤维化患者的肺成纤维细胞，每组分别施加或不施加TGFβ处理，每组设置3个生物学重复。