Processed Plaque-2.0 infection scores of the PCL screen for RV, imaged and analysed at EPFL

PCL compounds are indicated (name). The Plaque2.0-based infection scores of the four biological replicates of each PCL compound were averaged (mean). The Plaque2.0-based infection scores of the positive and negative controls are averaged (mean) over the 32 technical replicates, each, per plate. Each compound’s scores were normalized by the mean score of the negative control of the respective plate and indicated by Mean N_nuclei (number of nuclei based on Hoechst staining), Mean N_infected (number of GFP reporter-based infected nuclei), Mean InfIndex (ratio of numberOfInfectedNuclei to numberOfNuclei), Mean N_plaques (number of GFP reporter-based plaques) and Mean TotVirInt (total GFP reporter signal intensity). Non-toxic compounds were filtered by applying an inclusive μ- (mean of the negative control)  2σ (SD of the negative control) threshold for number of nuclei. Efficacy was filtered by applying an excluding μ-  2σ (SD of the negative control) threshold for the infection scores. The obtained scores for each infection score of each PCL compound indicated as Mean Scores N_nuclei (number of nuclei based on Hoechst staining), Scores N_Infected (number of GFP reporter-based number of infected nuclei), Scores InfIndex (ratio of numberOfInfectedNuclei to numberOfNuclei), Scores N_plaques (number of GFP reporter-based plaques) and Scores TotVirInt (total GFP reporter signal intensity). Compounds exhibiting significant toxicity to noninfected cells were excluded.

所有PCL化合物均以名称标注。针对每种PCL化合物，对其4次生物学重复的基于Plaque2.0的感染评分取均值。每块培养板的阳性、阴性对照的Plaque2.0感染评分，分别基于各自的32次技术重复取均值。各化合物的评分以对应培养板的阴性对照均值完成归一化处理，最终通过以下指标予以呈现：Mean N_nuclei（基于赫斯特（Hoechst）染色的细胞核总数）、Mean N_infected（基于绿色荧光蛋白（Green Fluorescent Protein, GFP）报告基因的感染细胞核数）、Mean InfIndex（感染细胞核数与总细胞核数的比值）、Mean N_plaques（基于GFP报告基因的噬斑数）以及Mean TotVirInt（GFP报告基因总信号强度）。针对细胞核数量指标，采用包容性阈值（阴性对照均值±2倍标准差，即μ±2σ）筛选无毒化合物。针对感染评分指标，采用排除性阈值（阴性对照均值±2倍标准差，即μ±2σ）筛选有效化合物。每种PCL化合物的各项感染评分最终以如下指标呈现：Scores N_nuclei（基于赫斯特（Hoechst）染色的细胞核总数）、Scores N_Infected（基于GFP报告基因的感染细胞核数）、Scores InfIndex（感染细胞核数与总细胞核数的比值）、Scores N_plaques（基于GFP报告基因的噬斑数）以及Scores TotVirInt（GFP报告基因总信号强度）。对未感染细胞表现出显著细胞毒性的化合物将被剔除。