Chromatin structure profiling of mouse muscle stem cells by ATAC-seq

By comparing ATAC-seq from freshly isolated satellite cells (QSC), satellite cells isolated 2.5 days post-injury (ASC), and satellite cells cultured for differentiation, we observed a gradual opening of satellite cell chromatin structure during activation and differentiation process. We used ATAC-seq to study the functional role of LncMyod during satellite cell differentiation and identified genes that exhibited decrease of accessibilities to their transcription start sites (TSS). The majority of the regions are myogenesis-related genes, suggesting LncMyod plays a critical role in establishing permissive chromatin environment for myogenic progression. Examination of chromatin accessibility using ATAC-seq in QSC, ASC, satellite cells cultured for differentiation with control siRNA (LncMyod Control), and satellite cells cultured for differentiation with siRNA targeting LncMyod (LncMyod Knockdown).

本研究通过对比新鲜分离的卫星细胞（QSC）、损伤后2.5天分离的卫星细胞（ASC）以及体外诱导分化培养的卫星细胞的ATAC-seq（转座酶可及性染色质测序）数据，观察到卫星细胞染色质结构在激活与分化过程中逐渐开放。我们利用ATAC-seq技术探究了LncMyod在卫星细胞分化过程中的功能，并筛选出转录起始位点（TSS）区域染色质可及性下降的基因。其中绝大多数差异区域对应肌发生相关基因，提示LncMyod在构建肌源性进展所需的允许性染色质微环境中发挥关键作用。本研究还通过ATAC-seq检测了以下四组样本的染色质可及性：QSC、ASC、转染对照siRNA的分化诱导卫星细胞（LncMyod对照组）以及转染靶向LncMyod的siRNA的分化诱导卫星细胞（LncMyod敲低组）。