Figure 4A

A. 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. 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. C. The spread of infection was quantified as the area of fluorescence using the NIS element software. D. 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. E. 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. F. The spread of infection was quantified as the area of fluorescence using the NIS element software.