Global gene expression profile of primary cultured keratinocytes with or without stimulation. Global gene expression profile of primary cultured keratinocytes with or without stimulation

Skin tissues from newborn C57BL/6 mice were obtained for primary keratinocyte culture. Cells were then stimulated with or without IL-25 (Biolegend) for 8hrs. Total RNAs were isolated with RNAprep pure Tissue Kit (DP431) and the RNA-seq library was prepared by the Beijing Genomics Institute. Sequence reads were obtained using BGIseq500 (Illumina) and successfully mapped to mouse genome. Reads counts were normalized based on RPKM, fold changes were calculated for all possible comparisons. Overall design: Gene expression primary cultured keratinocytes with or without stimulation

本研究采集新生C57BL/6小鼠的皮肤组织，用于原代角质形成细胞的体外培养。随后将培养得到的细胞分为两组，一组施加IL-25（Biolegend）刺激，另一组不做刺激处理，刺激时长均为8小时。采用RNAprep pure Tissue Kit（DP431）提取总RNA，并由北京基因组研究所完成RNA测序文库的构建。使用BGIseq500（Illumina）平台获取测序读段，所有读段均成功比对至小鼠参考基因组。对读段计数基于RPKM方法进行标准化，并针对所有可能的比较组计算基因表达的倍数变化。整体实验设计：对经IL-25刺激与未刺激的原代培养角质形成细胞开展基因表达分析。