Figure 8

Effect of eIF3L on yellow fever virus replication. (A) eIF3L overexpression in BHK21-rep-YFV17D LucNeoIres cells at 48 h post-transfection using an anti-Flag antibody or anti-GAPDH antibody as a control. eIF-pCDNAFlag, overexpressing eIF3L; pCDNAFlag, empty vector control; MOCK, control cells. (B) Luciferase activity at 48 h transfection with eIF3L in BHK21-rep-YFV17D LucNeoIres cell lysates; MOCK, control cells; pCDNAFlag, empty vector control; eIF3L-pCDNAFlag, eIF3L overexpression; siRNA NS1, positive control. *Significantly different between the RNAi NS1 and mock samples; **significantly different between the pCDNAFlag-eIF3L and mock samples. (C) Quantification of viral RNA transcripts at 24 and 48 h post-infection with pseudotyped lentiviral particles using real-time PCR analysis. pLKO.1-eIF3LshRNA1, eIF3L silenced and infected cells; pLKO.1-eIF3LshRNA2, eIF3L silenced and infected cells; pLKO, empty vector control-infected cells; non-target, plasmid control non-target infected cells; YFV, infected cells; MOCK, control cells. (D) The plaque reduction assay indicates a discrete reduction in viral replication. (E) Quantification of viral RNA in Vero cell supernatants overexpressing eIF3L at 6, 12, 24, and 48 h post-infection.