Figure 2

Immunofluorescence analysis of the expression of ZIKV E protein by MVA-ZIKV. (a) Detection of ZIKV E protein in the ER. HeLa cells were infected at 0.5 PFU/cell with MVA-ZIKV or MVA-WT for 24 h. Then, permeabilized cells were labeled with an anti-ZIKV E monoclonal mouse antibody and an anti-calnexin antibody. Anti-ZIKV E was detected with a mouse secondary antibody conjugated with the fluorochrome Alexa Fluor 488 (green). Anti-calnexin was detected with a rabbit secondary antibody conjugated with Alexa Fluor 594 (red). Cell nuclei were stained using DAPI (blue). The degree of co-localization of E and calnexin proteins is shown on the right by the yellow color. Scale bar: 10 μm. (b) Immunofluorescence analysis of ZIKV E protein in the cell membrane. HeLa cells were infected at 0.5 PFU/cell with MVA-ZIKV and at 24 h permebilized (upper panels) or non-permebilized (lower panels) fixed cells were stained with WGA probe conjugated to the fluorescent dye Alexa Fluor 594 (red) and a mouse monoclonal anti-ZIKV E antibody further detected with a mouse secondary antibody conjugated with the fluorochrome Alexa Fluor 488 (green). Cell nuclei were stained using DAPI (blue). Scale bars: 5 μm and 8 μm.