RNA sequencing analysis of IECs from WT and PIM1+/- mice

PIM1 is constitutively active and involved in various cellular processes, including cell survival, proliferation, and differentiation by phosphorylating a wide range of substrates. We investigated whether PIM1 is involved in the pathogenesis of ulcerative colitis (UC).We performed RNA sequencing analysis in intestinal epithelial cells (IECs) from PIM1-deficient mice and WT mice to study the molecular mechanisms through which PIM1 regulates goblet cell numbers. Gene expression profiles of IECs from WT and PIM1+/- mice were generated by deep sequencing, using Illumina NovaSeq 6000.To identify DEGs (differential expression genes) between two different samples, the expression level of each transcript was calculated according to the transcripts per million reads (TPM) method. RSEM was used to quantify gene abundances.PIM1为组成型激活蛋白，可通过磷酸化多种底物参与诸多细胞生物学过程，涵盖细胞存活、增殖与分化。本研究旨在探究PIM1是否参与溃疡性结肠炎（ulcerative colitis, UC）的发病机制。我们从PIM1缺陷小鼠与野生型（wild type, WT）小鼠中分离肠上皮细胞（intestinal epithelial cells, IECs），并对其开展RNA测序分析，以解析PIM1调控杯状细胞数量的分子机制。本研究通过深度测序构建了野生型与PIM1+/-小鼠肠上皮细胞的基因表达谱，测序平台采用Illumina NovaSeq 6000。为鉴定两组样本间的差异表达基因（differential expression genes, DEGs），本研究采用每百万转录本数（transcripts per million reads, TPM）法计算各转录本的表达水平，并使用RSEM对基因表达丰度进行定量。