Figure 4A

<b>A.</b> Lentivirus-mediated downregulation of MyD88. MRC-5 cells were transduced with control lentivirus or lentivirus encoding shRNA targeting MyD88 (shMyD88-1 and shMyD88-2) and selected using puromycin. Knockdown efficiency was assessed by western blot analysis. Blots were probed for MyD88 and p115 as loading control. <b>B. </b>The transduced and selected MRC-5 cells were infected with TB40/E-mCh virus at MOI of 0.05 and spread of the virus was monitored by confocal imaging. Representative images from day 3, 9 and 15 are shown. <b>C. </b>The spread of infection was quantified as the area of fluorescence using the NIS element software. <b>D. </b>MRC-5 cells were transduced with lentivirus from control or two different cassettes encoding the MyD88 gene (MyD88-1 and MyD88-2) and subjected to puromycin selection. Cell lysates were harvested and run on western blot to analyse MyD88 levels. Tubulin served as a loading control. <b>E. </b>The transduced and selected MRC-5 cells were infected with TB40/E-mCh virus at MOI of 0.05 and the spread of the virus was monitored every three days by confocal imaging up to day 21. Representative images from day 3, 9 and 15 are shown. <b>F. </b>The spread of infection was quantified as the area of fluorescence using the NIS element software.

<b>A.</b> 慢病毒介导的髓样分化因子88（MyD88）下调。将MRC-5细胞用对照慢病毒或编码靶向MyD88的短发夹RNA（short hairpin RNA，shRNA）的慢病毒（shMyD88-1和shMyD88-2）进行转导，并以嘌呤霉素进行筛选。通过蛋白质印迹法（western blot）评估敲低效率，以MyD88和作为内参的p115蛋白为检测靶标。<b>B.</b> 对经转导及筛选的MRC-5细胞以感染复数（Multiplicity of Infection，MOI）0.05感染TB40/E-mCh病毒，通过共聚焦成像监测病毒扩散情况，展示第3天、第9天及第15天的代表性图像。<b>C.</b> 采用NIS Elements软件以荧光面积为指标对感染扩散程度进行定量分析。<b>D.</b> 将MRC-5细胞用对照慢病毒或两种编码MyD88基因的不同表达盒的慢病毒（MyD88-1和MyD88-2）进行转导，并经嘌呤霉素筛选。收集细胞裂解液进行蛋白质印迹分析，以检测MyD88的表达水平，以微管蛋白（Tubulin）作为内参。<b>E.</b> 对经转导及筛选的MRC-5细胞以感染复数（MOI）0.05感染TB40/E-mCh病毒，每3天通过共聚焦成像监测病毒扩散情况直至第21天，展示第3天、第9天及第15天的代表性图像。<b>F.</b> 采用NIS Elements软件以荧光面积为指标对感染扩散程度进行定量分析。