Effect of time of addition of compounds on NiV antiviral activity for brilliant green (a) gentian violet (b) and gliotoxin (c). Inhibition of NiV infection as measured by viral nucleoprotein expression determined by immunoassay. For cell pre-incubation (Cells), each compound was incubated with cells for 60 min, media was then removed and virus inoculum (1,000 TCID50/ml NiV) added and incubated for 60 min, then inoculum was removed and replaced with EMEM-10 and cells were incubated overnight. For Pre-infection (Pre), virus was incubated with compound for 60 min, then added to cells for 60 min prior to replacement with EMEM-10 overnight. Co-infection (Co) wells received compound and virus inoculum simultaneously, incubated for 60 min, then media was replaced with EMEM-10 and cells were incubated overnight. Post-infection (Post) wells were infected with virus for 60 min followed by inoculum removal and replacement with compound in media. Cells were incubated and assayed as above. Values are expressed as the Mean +/- S.E (n = 3).