: Proximity proteomics identifies endothelial cell septins and PAK2 as decisive regulators of actomyosin mediated expulsion of von Willebrand factor.

A Rab27a targeted APEX2 construct was transfected into primary human umbilical endothelial cells (HUVEC) and the proximal proteins surrounding endothelial storage organelles (Weibel Palade bodies) identified by proximity proteomics in unstimulated and stimulated conditions. The procedure was carried out in the presence and absence of two different stimuli (10 minutes-phorbol myristate acetate or a combination of Histamine, adrenalin and 3-isobutyl-1-methylxanthine) and the data presented represents 4 independent experiments each with 2 technical replicates.

将靶向Rab27a的APEX2融合表达载体转染至原代人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVEC），随后通过邻近蛋白质组学（proximity proteomics）技术，分别在未刺激与刺激条件下鉴定内皮储存细胞器——魏贝尔-帕拉德小体（Weibel-Palade bodies）周围的邻近蛋白。本实验设置了两种不同的刺激方案，分别为以十四烷酰佛波醇乙酸酯（phorbol myristate acetate）刺激10分钟，以及以组胺（Histamine）、肾上腺素（adrenalin）与3-异丁基-1-甲基黄嘌呤（3-isobutyl-1-methylxanthine）联合刺激，同时设置未刺激对照组；本次公开的数据集包含4次独立实验，每次实验均设置2个技术重复。