Fig 2.

Productive infection of LACV is dependent on dynamin- and clathrin-mediated endocytosis. BHK-21 cells (untreated or pretreated with DMSO, DYN, CPZ, or sucrose) were mock infected or infected with LACV in the presence of each treatment for 1 h. (A) To assess the effect of DYN, CPZ, and sucrose on LACV replication, cells were infected with LACV (MOI, 0.005) and supernatants were harvested 24 h postinfection. Viral titers were determined by standard plaque assays on VERO cells. The results are means ± SD of 3 independent experiments (*, P < 0.01 by Tukey test). (B) A fusion-from-within (FFWI) assay was used to measure the extent of LACV infection 18 h postinfection (MOI, 0.5). FFWI in LACV-infected cells was significantly decreased by inhibitors of dynamin- and clathrin-mediated endocytosis. As expected, cell-to-cell fusion was not observed in mock-infected control cells (data not shown) and LACV-infected cells fused at 4°C. Error bars represent standard errors of the means from three independent experiments performed in duplicate (*, P ≤ 0.01; **, P ≤ 0.001; both by Tukey test). The dashed line represents the limit of detection.