Respiratory syncytial virus infection
Primary NECs grown at ALI were infected with green fluorescent tagged RSV-A2 at a multiplicity of infection (MOI) of 1 plaque-forming unit (pfu) per epithelial cell. Cells were incubated with virus for 2 hours in PBS at 37oC in the apical chamber, after which they were washed with PBS and TEER recorded. RSV load was quantified (pfu/ml) using a plaque assay in confluent monolayers of Hep-2 cells. Cultures were maintained in hydrocortisone free media for 6 days before wounding. Apical and basal wash was collected at 6 days to determine viral titre and toxicity of the virus.