FIG 8.

Passaging in the presence of ALV reduces EAV sensitivity toward both ALV and CsA. Virus was passaged at increasing ALV concentrations, ranging from 4 μM during passage 1 (P1) to 75 μM during P10. P10 harvests of EAV replicates ALV-1 and ALV-2 were tested for resistance to ALV (A) and CsA (B) treatment. Huh7 cells in 96-well plates were infected at an MOI of 0.05 in the presence of 0 to 50 μM ALV or CsA. Cells were incubated for 3 days, and cell viability was monitored using a commercial assay. In addition, the cytotoxicity of CsA treatment was monitored in parallel in mock-infected Huh7 cells. Graphs show the results (average and SD) of a representative experiment that was performed in quadruplicate. All experiments were repeated at least twice.