Fig 2.

Identification of TIM-1 IgV residues that impact EBOV GP-dependent entry. (A to C) Relative virus transduction into HEK 293T cells mediated by mutant TIM-1 constructs compared to that with WT TIM-1. At 48 h, transfected cells were transduced with EBOV GP-VSVΔG (A), LASV GPC-VSVΔG (B), MARV GP-VSVΔG (C), or full-length EBOV-VSVΔG (C) pseudovirions. (A and B) Transductions were also done in the presence or absence of anti-human TIM-1 MAb ARD5 (1.7 μg/ml). EGFP expression was assessed 24 h later. (D) EBOV GP/rVSV-EGFP infection mediated by TIM-1 constructs in HEK 293T cells (MOI of 1 as determined from titers in Vero cells) relative to WT TIM-1. (E) Serial dilution of EBOV GP/rVSV-EGFP on TIM-1-transfected HEK293T cells. EGFP expression was assessed 24 (A to C) or 48 (D and E) h later by flow cytometry. Data are shown as means ± SEM for at least three replicates. (A to D) Significance was calculated using one-sample t-test comparison to 100 (**, P < 0.001; *, P < 0.01). Significance for all ARD5 transductions for EBOV was defined as P < 0.001.