Additional file 1 of Endomucin selectively regulates vascular endothelial growth factor receptor-2 endocytosis through its interaction with AP2

Additional file 1: Supplemental Figure 1. Markov Clustering of EMCN-binding proteins identified by mass spectrometry. (A) EMCN-binding proteins identified through mass spectrometry were analyzed and visualized using the STRING database (https://string-db.org/). The network type was set to physical subnetwork, and the significance of network edges was based on confidence, with a minimum required interaction score of high confidence (0.700). Further functional clustering of EMCN-binding proteins was performed using Markov clustering. Proteins within the same cluster were connected by solid lines and marked with the same color, while dashed lines indicated boundaries between clusters. (B) The table lists the top five protein clusters with the highest number of proteins. Supplemental Figure 2. Confirmation of EMCN knockdown. (A) Representative image of the western blot assay showing significant reduction of EMCN at protein level in HRECs transfected with siEMCN compared to siNT control. (B) Quantification of EMCN protein by western blot analysis demonstrates a significant decrease of EMCN at 72 hr after transfection. Student-t test was used. ****P<0.0001, n=3. (C) Quantification of EMCN mRNA indicates a significant decrease of EMCN (1.024 ± 0.126 vs. 0.0128 ± 0.0003) at 48 hr after transfection. Student-t test was used. ****P<0.0001, n=3.

附加文件1：补充图1。质谱（mass spectrometry）鉴定的EMCN结合蛋白的马尔可夫聚类（Markov Clustering）。(A) 采用STRING数据库（STRING database，https://string-db.org/）对通过质谱鉴定得到的EMCN结合蛋白进行分析与可视化。将网络类型设置为物理子网，网络边的显著性基于置信度，最低要求相互作用评分设为高置信度（0.700）。进一步采用马尔可夫聚类对EMCN结合蛋白进行功能聚类。同一聚类内的蛋白以实线连接并使用相同颜色标记，虚线则表示聚类间的边界。(B) 该表格列出了蛋白数量最多的前五大蛋白聚类。补充图2：EMCN敲低验证。(A) 蛋白质免疫印迹（western blot）试验的代表性结果显示，与siNT对照组相比，转染siEMCN的人类视网膜内皮细胞（HRECs）中EMCN的蛋白水平显著降低。(B) 蛋白质免疫印迹定量分析结果表明，转染后72小时EMCN蛋白水平显著下降，采用学生t检验（Student-t test），****P<0.0001，n=3。(C) EMCN mRNA定量结果显示，转染后48小时EMCN mRNA水平显著下降（1.024 ± 0.126 相较于 0.0128 ± 0.0003），采用学生t检验（Student-t test），****P<0.0001，n=3。