Fig. 5.

The effects of a cathepsin L inhibitor or chloroquine in PEC entry into the cells. Confluent LLC-PK cells were pre-treated with mock (medium), Z-FY-CHO or chloroquine for 30 min before virus inoculation. Cells were then inoculated with PEC (MOI 50) in the presence of mock, Z-FY-CHO or chloroquine. GCDCA (100 µM) was present in cell culture during virus infection to support PEC replication, with the exception of mock-infected cells and PEC-infected negative control cells (PEC w/o GCDCA) . Following virus infection for 1 h, cells were fixed and probed with rabbit polyclonal anti-Rab7 or swine anti-PEC polyclonal antibodies, followed by secondary antibodies of PerCP-Cy5.5 labeled secondary antibody against Rab7 (red, k–o) or FITC-labeled secondary antibody against PEC (green, f–j). Nuclei were stained with sytox orange (pseudo colored blue, a–e). Confocal images on the prepared samples were obtained and colocalization analysis of PEC and Rab7 was performed using ImageJ software. In the merged images (p–t), colocalization of PEC (green) and Rab7 (red) appears in white color.