FIG 4.

MAbs c2G4 and c4G7 do not inhibit trafficking of EBOV GP VLPs to NPC1⁺ endolysosomes. VLPs bearing EBOV-Mayinga GP were pretreated with 50 μg/ml c2G4 or c4G7, added to BSC1 cells, and processed for VLP trafficking and colocalization with NPC1 as described in Materials and Methods. Control cells had been preincubated with 20 μM nocodazole (Noc) or DMSO (Mock) for 1 h prior. (A) Data are the average Manders coefficients for colocalization of mCherry-VLPs (red) with NPC1 (stained green) from 27 (Mock and Noc treated) or 33 (c2G4 and c4G7 treated) microscope fields. Error bars represent the SD. ***, P < 10⁻¹⁵ (relative to mock samples). (B) Representative images are shown. White arrows indicate areas of VLP-NPC1 colocalization, and red arrows indicate VLPs that have not colocalized with NPC1. Scale bars, 10 μm (primary images) and 2 μm (insets). Virtually identical results were seen in two additional experiments.