Comparison of thymic mTEC cells from MyD88fl/fl x Foxn1Cre and MyD88fl/fl mice by transcription profiling

We compared in vitro mTECs-high from MyD88fl/fl x Foxn1Cre and MyD88fl/fl mice: Thymi were enzymaticaly digested, cells were MACS enriched for CD45- fraction, and FACS sorted using BD Influx sorter. mTECs-high were gated as EpCAM+CD11c-Ly51-MHCII+CD80+. Total RNA was isolated using RNeasy Plus Micro Kit (Qiagen). 5 samples per condition were used.我们对来自MyD88fl/fl × Foxn1Cre小鼠与MyD88fl/fl小鼠的体外高表达型髓质胸腺上皮细胞（medullary thymic epithelial cells-high, mTECs-high）进行了对比分析。具体实验流程如下：首先通过酶解法消化小鼠胸腺组织，随后采用磁激活细胞分选（Magnetic Activated Cell Sorting, MACS）富集CD45阴性细胞组分，再使用BD Influx流式细胞分选仪完成荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）。高表达型髓质胸腺上皮细胞的分选门控策略为EpCAM+CD11c-Ly51-MHCII+CD80+。总RNA提取采用RNeasy Plus Micro试剂盒（Qiagen），每个实验条件设置5个生物学重复样本。