Salmonella effector SopB reorganizes cytoskeletal vimentin to maintain replication vacuoles for efficient infection

The raw date of high-content imaging based screening. The specific screening process is as follows: Actin-mCherry and vimentin-GFP double-labeled stable expression cells were seeded in 384-well plates. After cells reached 70% - 80% confluence, 1694 inhibitors from Selleck chemicals library (US) were added at a dose of 1 µM, respectively. Three duplicates were performed for each compound. After 12 hours of treatment, cells were washed three times with PBS and fixed with 4% PFA. The nuclei were counterstained with DAPI, and cells were imaged by Operetta CLS High-content Imaging and Analysis System (PerkinElmer, US).

本数据集为基于高内涵成像（high-content imaging）的筛选实验原始数据。具体筛选流程如下：将肌动蛋白-mCherry（Actin-mCherry）与波形蛋白-GFP（vimentin-GFP）双标记的稳定表达细胞接种于384孔板中。待细胞汇合度达到70%~80%时，分别以1 µM的浓度加入来自美国Selleck化学品文库的1694种抑制剂。每种化合物设置三次重复实验。处理12小时后，用磷酸盐缓冲液（Phosphate Buffered Saline, PBS）洗涤细胞三次，再以4%多聚甲醛（Paraformaldehyde, PFA）固定细胞。随后用4',6-二脒基-2-苯基吲哚（DAPI）对细胞核进行复染，并通过Operetta CLS高内涵成像分析系统（PerkinElmer, 美国）完成细胞成像。