West Nile Virus Replication Requires Fatty Acid Synthesis but Is Independent on Phosphatidylinositol-4-Phosphate Lipids
Figure 1
Analysis of cellular components involved in WNV replication complex.
(A) Ultrastructure of WNV-induced membrane alterations. Cells infected with WNV (MOI of 5 PFU/cell) were fixed and processed for electron microscopy at 24 h p.i. (a) Electron micrograph showing membrane alteration on WNV infected cells: convoluted membranes (CM), WNV induced-vesicles (Ve), vesicle packets (VP), and electron dense virions (Vi). (b) Higher magnification images of VP induced by WNV infection. Black arrows indicate the point of contact between a vesicle and the outer membrane of the VP. Asterisks denote Ve with electron dense fibrous material. (c) WNV virions trafficking through the Golgi complex. (d) Whorls of stacked membranes. Scale bars: 200 nm. (B) Cells infected or not (mock) as in (A) were fixed and processed for immunofluorescence and confocal microscopy. WNV dsRNA was detected using a monoclonal antibody and cellular structures were labelled by using specific antibodies, or by transfection with plasmids encoding fluorescent fusion proteins (see text for details). Suitable secondary antibodies coupled to AF488 or 594 were used. Scale bar: 10 µm.
