Enforced MAB21L4 expression enhances epidermal diferentiation

RNA-seq was used to define the role of MAB21L4 in epidermal differentiation by placing primary human keratinocytes transduced to overexpress MAB21L4 onto human dermis and grown for three days at the air-liquid interface to generate organotypic skin tissue. Total RNA was then isolated from human skin organoids using QIAshredder (Qiagen) and Trizol (ThermoFisher) followed by DNA removal with the TURBO DNA-free kit (Ambion) according to the manufacturer’s instructions. RNA integrity was verified using an Agilent 2100 Bioanalyzer. RNA-Seq libraries were prepared with the mRNA Seq Sample Prep Kit (Illumina, Inc.) as recommended by the manufacturer. 100 bp paired-end sequencing reads were obtained using the Illumina HiSeq platform.

本研究采用RNA测序（RNA-seq）技术，通过将经转导以过表达MAB21L4的原代人角质形成细胞接种于人真皮，并在气液界面培养3天以构建器官型皮肤组织，以此明确MAB21L4在表皮分化中的作用。随后，依照制造商说明书，使用QIAshredder（Qiagen）与Trizol（ThermoFisher）从人皮肤类器官中提取总RNA，并采用TURBO DNA-free试剂盒（Ambion）去除基因组DNA残留。采用安捷伦2100生物分析仪（Agilent 2100 Bioanalyzer）验证RNA完整性。参照制造商推荐的操作规范，使用mRNA测序样本制备试剂盒（Illumina, Inc.）构建RNA-seq文库。通过Illumina HiSeq平台获取100 bp双端测序读段。