Figure 4. Establishment of persistent infection in Ae. albopictus cell lines.

A. Titres of virus released into the supernatant from each passage of cells. Cells were initially infected at MOI of 0.1 PFU/cell, and passaged regularly at a 1∶5 split ratio. The supernatant medium was collected at the time of each passage and the presence of virus measured by plaque assay. B. Detection of viral N protein. An aliquot of cells was collected at each passage, lysed and 10 µg of total protein was analyzed by Western blotting with anti-BUNV N protein antibodies. Numbers at the top of the blots indicate the passage number.