Figure 1

An assay for the collection of secreted proteins from JEV-infected BHK-21 cells. A) A flowchart outlining the protocol for the identification of JE virus replication from intracellular lysates and secreted proteins from the JEV-infected cells. B) Effect of serum deprivation on JEV replication. Cells were incubated with medium supplemented with 2% FBS or serum-free medium for 24 hours and viral proteins, NS1 and NS5, were assessed by Western blot analysis using anti-NS5/anti-NS1 polyclonal antibodies. Extracts from mock-infected cells serve as a negative control. Two independent replicates are shown for the serum-free and 2% serum conditions. C) Effect of serum deprivation on cell viability. β-actin was not detected in the secreted medium of mock- and JEV-infected BHK-21 cells. Three independent replicates are shown for the mock- and JEV-infected conditions.