Fig. 5. N-linked glycans on EBOV GP prevent activation of CDC mechanism.

a SNAP-tagged 293F cells expressing EGFP and EBOV GP were treated with the vehicle control (–) or 1 µg ml⁻¹ tunicamycin (+) and incubated with 10 µg ml⁻¹ mAbs. Binding of mAbs to EBOV GP was determined by flow cytometry using PE-conjugated goat anti-human IgG secondary antibody. b SNAP-tagged 293F cells were treated as in (a), and the surface expression of EBOV GP was determined by flow cytometry using rabbit anti-EBOV VLP antiserum and mouse anti-rabbit IgG secondary antibody conjugated with PerCP-Cy5.5. c Cells were treated as in (a), and CDC assay was performed as in Fig. 1c. The percentages of EGFP^–AF647⁺ cells in samples treated with the vehicle control or tunicamycin and incubated with DENV-2D22 mAb were used for background signal subtraction. Mean ± SEM of triplicate samples are shown. *p < 0.01; **p < 0.001; ***p < 0.0001; ns, not significant (unpaired t-test). Representative flow cytometry dot plots are shown.