FIG. 3.

Dynamin is necessary for efficient ZEBOV-GP pseudovirion entry into Axl-dependent cells. (A to C) Ability of DN dynamin to inhibit FIV-ZEBOVΔO transduction. SNB19 cells (A), Hffs (B), and HuVECs (C) were transduced with adenoviral vectors bearing GFP (Ad-GFP) or a dominant negative form of dynamin 2 (Ad-DN-dyn2) at an MOI of 30 for SNB19 cells and an MOI of 90 for Hffs and HuVECs, ensuring greater than 90% adenovirus transduction. Eighteen hours following adenoviral transduction, SNB19 (A) and Hff (B) cells were incubated with FITC-labeled dextran (0.5-mg/ml final concentration) for 1 h or transduced with FIV pseudovirions (MOI, 0.005; all cell populations) for 48 h. SNB19 and Hff cells were analyzed by flow cytometry for uptake of dextran after 1 h. Transduced cells were fixed and stained for β-Gal activity after 48 h. (D) Ability of Dynasore to inhibit FIV-ZEBOVΔO transduction. SNB19 cells were treated for 1 h with the indicated amounts of Dynasore. Treated cells were incubated with FITC-conjugated dextran or Cy5-conjugated Tfr or CTb for 1 h and analyzed by flow cytometry or incubated with FIV pseudovirions (MOI, 0.005) for an additional 6 h in the presence of the drug. The medium on cells transduced with FIV-ZEBOVΔO or FIV-VSV-G was refreshed after 6 h with medium not containing drug, and transduced cells were fixed and stained for β-Gal activity 48 h following transduction. Cell viability in the presence of the treatments is shown as a dashed line in all panels. Data represent the averages and standard errors of three experiments performed in triplicate. *, P < 0.05; **, P < 0.001.