Mock infected controls for transcriptional profiling of infections with high capacity MCMV vector.

To gain a detailed knowledge about the virus cycle of murine cytomegalovirus (MCMV) and its high capacity vector in cross-species settings, we analyzed the early and late viral and host transcriptome upon infection of human cells. ARPE-19, A549, and 911 cells (5.0×105 cells/well) were infected in a 24-well format 4 h post-seeding with MCMV-wt or Q4-LRBAs-GLuc, a high capacity replication competent vector based on MCMV, at an MOI of 3. As control, we used infection of mouse embryonic fibroblast, which is the natural host of the wild type MCMV, treated similarly. Harvesting occurred at 8 hpi and 31 hpi through centrifugation at 1.000 g for 5 min. Cell pellets were washed with PBS, re-suspended in 350 µL of RLT buffer, and processed using the RNeasy Mini kit as per the manufacturer’s instructions in independent triplicates. At least 1.500 ng of RNA in a 30 µL volume was isolated and sent for Illumina next-generation sequencing, resulting in paired-end sequencing with a read length of 2×100 bp and a depth of 20 million reads. This submission contains the reads, we obtained analyzing the non-infected cell lines.为深入解析鼠巨细胞病毒（murine cytomegalovirus, MCMV）的病毒生命周期及其跨物种感染场景中的高效载体潜能，我们对人类细胞感染后的早期与晚期病毒及宿主转录组展开了系统分析。我们将ARPE-19、A549及911细胞（每孔5.0×10⁵个细胞）在铺板4小时后，以感染复数（multiplicity of infection, MOI）为3的感染比例，使用野生型MCMV（MCMV-wt）或基于MCMV构建的高效复制型载体Q4-LRBAs-GLuc，在24孔板中进行感染。作为对照，我们以相同处理流程感染了野生型MCMV的天然宿主——小鼠胚胎成纤维细胞。分别于感染后8小时（hours post infection, hpi）与31小时（hpi），通过1000 g离心5分钟收集细胞。收集得到的细胞沉淀经磷酸盐缓冲液（phosphate buffered saline, PBS）洗涤后，重悬于350 μL RLT缓冲液中，并依照制造商说明书使用RNeasy Mini试剂盒完成独立三次重复的RNA提取。最终从30 μL体系中分离得到至少1500 ng总RNA，随后送至Illumina平台进行下一代测序，采用读长为2×100 bp的双端测序策略，测序深度达2000万条reads。本提交包含我们在分析未感染细胞系过程中获取的reads数据。