Figure 2

Ebola GP lacking the mucin-like region does not accumulate in the ER. A. HEK293T cells were transiently transfected for 24 hours with 8 μg GPΔmucin (pCDNA6-EbGPΔmucin-mutΔ1234) and 2 μg pDsRed2-ER vector (Clontech) that labels the endoplasmic reticulum (ER, red). Cells were fixed and stained for NPC proteins (blue) using mouse monoclonal antibody 414 (Covance Research Products) and GPΔmucin (green) using the KZ52 neutralizing human monoclonal antibody labeled with a Zenon labeling kit (Molecular Probes). Scale bar represents 15 μm. Side panels show individual fluorescent channels from the boxed region in the image. B. HEK293T cells were transiently transfected for 24 hours with 8 μg GPΔmucin and 2 μg pEYFP-Golgi vector (Clontech) that labels the trans-medial region of the Golgi apparatus (blue). Cells were fixed and stained for early endosomes using a mouse monoclonal antibody against EEA1 (BD Biosciences, green) and for GPΔmucin as above (red). Scale bar represents 15 μm. Side panels show individual fluorescent channels from the boxed region in the image. C. HEK293T cells were transiently transfected with 10 μg GPΔmucin for 24 hours. Cells were fixed and stained for late endosomes using a mouse monoclonal antibody targeting CD63 (BD Biosciences, blue) and for GPΔmucin as above (red). Scale bar represents 15 μm. Side panels show individual fluorescent channels from the boxed region in the image.