m6A-seq of Min6 cells were transfected with the GR cDNA expression vector or control vector. m6A-seq of Min6 cells were transfected with the GR cDNA expression vector or control vector

To further explore the underlying molecular mechanism of GR’s effects, we transfected the Min6 cell with the GR cDNA expression vector or control vector, and then extracted RNA of Min6 cells. M6A antibody was used to specifically recognize m6A on mRNA. methylation modified RNA fragments were obtained by immunoprecipitation and used for high-throughput sequencing. By comparing the captured RNA fragments, we found that there were 22381 difference peaks between GR overexpression group and control group, among which 3659 binding peaks were significantly different.Further analysis the MeRIP sequencing results of MIN6 cells overexpressing GR revealed that decreased m6A was detected in a large number of autophagy-related mRNAs. Overall design: Input and m6A-IP mRNA of Min6 cells transfected Control vector or GR expression vector

为进一步探究糖皮质激素受体（Glucocorticoid Receptor, GR）发挥作用的潜在分子机制，我们将GR cDNA表达载体或空对照载体转染至Min6细胞中，随后提取Min6细胞的RNA。使用m6A特异性抗体靶向识别mRNA上的N6-甲基腺苷（m6A）修饰，通过免疫沉淀获取携带甲基化修饰的RNA片段，并将其用于高通量测序。通过比对所捕获的RNA片段，我们发现GR过表达组与对照组间共存在22381个差异峰，其中3659个结合峰存在显著差异。对过表达GR的MIN6细胞的MeRIP测序结果开展进一步分析后发现，大量自噬相关mRNA的m6A修饰水平显著下调。实验设计概述：转染空对照载体或GR表达载体的Min6细胞的Input mRNA及m6A免疫沉淀（IP）mRNA。