Fig 1. Quantification of SEOV infection in human and rat endothelial cells.

A) Representative images of mock or SEOV-infected RLMVEC, HUVEC, and Vero E6 cells (1,8 dilution) at 10x magnification. Nuclei are stained with DAPI (blue) and SEOV N protein is shown in green. B) Percentage of SEOV N-positive cells at each 1:2 viral dilution from neat to 1:1024 dilution and mock to determine background fluorescence, quantified using the CellInsight CX7 imaging platform for all cell types. C) Calculated infectious units/mL for each cell type based on focus-forming unit (FFU) assay for Vero E6 and CellInsight CX7 imaging for Vero E6, RLMVEC and HUVEC (mean ± SD, from four replicate wells per dilution, across three independent experiments). D) Percentage of SEOV N-positive RLMVEC over five days, following infection with MOIs of 0.05, 0.1, and 0.25, quantified using CellInsight CX7. E) Representative images of SEOV-infected RLMVEC at each MOI and timepoint, captured at 20x magnification.