Single-cell RNA-seq analysis of WT and Furin KO TECs. Single-cell RNA-seq analysis of WT and Furin KO TECs

To assess the effect of Furin ablation on TEC development and heterogeneity, we detected the transcriptional profiles of 2-week-old WT and Furin KO TECs using single-cell RNA-seq. Employing an unsupervised graph-based clustering strategy, 20 clusters of TECs and 5 clusters of non-TEC contaminants were identified. After removing contaminants, 4301 WT TECs and 4086 Furin KO TECs were obtained. UMAP visualization revealed that the proportion of mTECs decreased significantly and the proportion of cTECs increased significantly in TEC-specific Furin knockout mice in comparison with WT mice. Further analysis showed that Furin ablation did not affect the proportion of mTEC I, II and III subsets, but reduced the proportion of mTEC IV subsets markedly. Overall design: Compare the transcriptional profiles of 2-week-old WT and Furin KO TECs using single-cell RNA-seq

为评估弗林蛋白酶（Furin）敲除对胸腺上皮细胞（TEC, Thymic Epithelial Cell）发育与异质性的影响，我们借助单细胞RNA测序（single-cell RNA-seq）检测了2周龄野生型（WT）与弗林蛋白酶敲除（Furin KO）胸腺上皮细胞的转录组谱。本研究采用无监督图基聚类策略，共鉴定出20个胸腺上皮细胞簇与5个非胸腺上皮细胞污染簇。去除污染细胞后，最终获得4301个野生型胸腺上皮细胞与4086个弗林蛋白酶敲除胸腺上皮细胞。均匀流形近似与投影（UMAP, Uniform Manifold Approximation and Projection）可视化结果显示，相较于野生型小鼠，胸腺上皮细胞特异性弗林蛋白酶敲除小鼠的髓系胸腺上皮细胞（mTEC, Medullary Thymic Epithelial Cell）占比显著降低，而皮质胸腺上皮细胞（cTEC, Cortical Thymic Epithelial Cell）占比显著升高。后续深入分析表明，弗林蛋白酶敲除未对髓系胸腺上皮细胞I、II、III亚群的占比产生影响，但显著降低了髓系胸腺上皮细胞IV亚群的占比。实验整体设计：采用单细胞RNA测序技术，对比2周龄野生型与弗林蛋白酶敲除胸腺上皮细胞的转录组谱。