Figure 3

Co-expression of Ebola GP, VP40 and NP in HEK293T cells does not alter localization of individual proteins. A. HEK293T cells were co-transfected with 5 μg Ebola GP and 5 μg eGFP-VP40 plasmids for 24 hours. Cells were fixed and stained for NPC proteins using mouse monoclonal antibody 414 (Covance Research Products, blue) and GP (red) using KZ52 neutralizing human monoclonal antibody labeled with a Zenon labeling kit (Molecular Probes). Green represents VP40. Scale bar represents 15 μm. Upper and lower side panels show VP40 and GP fluorescence respectively, from the boxed region in the image. B. HEK293T cells were transfected with 10 μg Ebola NP plasmid for 24 hours. Cells were fixed and stained for DNA using Hoechst (blue), NP using a mouse monoclonal antibody targeting NP (red) and vimentin using a mouse monoclonal antibody targeting vimentin (green). Scale bar represents 5 μm. Upper and lower side panels show the vimentin and DNA fluorescence and the NP and DNA fluorescence respectively, from the boxed region in the image. C. HEK293T cells were co-transfected with 5 μg Ebola NP and 5 μg eGFP-VP40 plasmids for 24 hours. Cells were fixed and stained for NP using a mouse monoclonal antibody targeting NP (red) and DNA using Hoechst (blue). Green represents VP40. Scale bar represents 5 μm. Upper and lower side panels show the VP40 and DNA fluorescence and the NP and DNA fluorescence respectively. D. HEK293T cells were co-transfected with 5 μg Ebola NP and 5 μg eGFP-VP40 plasmids for 24 hours. Cells were fixed and stained for NP using a mouse monoclonal antibody targeting NP (blue) and filamentous actin using Texas Red Phalloidin (Invitrogen, red). Green represents VP40. Scale bar represents 15 μm. Side panels show the individual fluorescent channels from the boxed region in the image.