Processed Plaque-2.0 infection scores of the PCL screen, 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-  3σ (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.

PCL化合物已标注其名称。针对每种PCL化合物的4次生物学重复，基于Plaque2.0平台的感染评分取均值。阳性对照与阴性对照基于Plaque2.0平台的感染评分，在每块平板的32次技术重复中分别取均值。每种化合物的评分均以对应平板的阴性对照均值进行归一化，相关指标包括：基于赫斯特染色（Hoechst staining）的细胞核均值计数（Mean N_nuclei）、基于绿色荧光蛋白（GFP）报告系统的感染细胞核均值计数（Mean N_infected）、感染指数均值（Mean InfIndex，即感染细胞核数与细胞核总数的比值）、基于GFP报告系统的噬斑均值计数（Mean N_plaques）以及总GFP报告信号强度均值（Mean TotVirInt）。通过对细胞核计数设置阴性对照均值（μ）±2倍标准差（SD）的包容性阈值，筛选出无细胞毒性的化合物；通过对感染评分设置阴性对照均值（μ）±3倍标准差（SD）的排他性阈值，筛选出具有药效活性的化合物。针对得到的每种PCL化合物的各项感染评分，分别标注为：评分细胞核数（Scores N_nuclei，基于赫斯特染色的细胞核总数）、评分感染细胞核数（Scores N_Infected，基于GFP报告系统的感染细胞核总数）、评分感染指数（Scores InfIndex，即感染细胞核数与细胞核总数的比值）、评分噬斑数（Scores N_plaques，基于GFP报告系统的噬斑数）以及评分总GFP报告信号强度（Scores TotVirInt，总GFP报告信号强度）。最终将对未感染细胞表现出显著毒性的化合物予以排除。