Figure 4.

Weak MojV F and G-mediated cell–cell fusion is detected with cells expressing human and rodent A-class ephrin ligands. Cell–cell fusion reporter assays were performed as described in Figure 3 with the following modifications. CHO-K1 target cells were co-transfected with the pTM1-Nluc reporter plasmid and one ephrin-expressing plasmid at a time or an empty vector (pcDNA). Cell mixtures of effector and target cells were assayed for Nano luciferase activity. (A) CedV F and G or (B) MojV F and G effector cells were tested for their ability to mediate cell–cell fusion against a panel of target cells transiently expressing human and rodent ephrin ligands. Maximum RLUs were recorded 48h post-application of target cells to effector monolayers co-expressing CedV F and G constructs. Co-cultures containing effector cells co-transfected with MojV F and G constructs yielded maximum RLUs on day 6. The grey line represents the normalized reference level of 1 (background). The experiments were performed at least 3 times in technical triplicates. m: mouse ephrin ligand; h: human ephrin ligand; r: rat ephrin ligand. Data were analyzed by Welch’s t test comparing the normalized maximum RLU means of F and G expressing effector cells co-cultured with target cells transfected with an ephrin construct to effector cells co-cultured with target cells transfected with an empty vector (pcDNA). The error bars represent the standard deviation. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.