WNV replication kinetics in Se deficient cells. (A) Vero cells grown in control, Se-, and Se+ media were infected with WNV at day 7 post induction of Se deficiency and cell supernatants were harvested every 24 hr for 5 days. Viral RNA extracted from the cell supernatant was used to determine viral copy number by qRT-PCR and expressed as viral copy number per mL. Data represents mean ± SD of three independent infections. (B) WNV-infected control, Se- and Se+ Vero cells grown and fixed on coverslips at day 2 post infection were incubated with monoclonal human anti-WNV env antibody and then with Alexa Fluor 488 conjugated goat anti-mouse secondary antibody. Mock infected Vero cells (a) and infected Vero cells stained with secondary antibody alone (b), were used as a negative control. The experiments were performed in triplicate and c, d and e represents WNV antigen staining in Vero cells grown in control, Se- deficient and Se- adequate media, respectively. Scale bar represents 10 μm at a magnification of 63× in all pictures.