2.2. Propagation of ZIKV

ZIKV strain ({"type":"entrez-nucleotide","attrs":{"text":"KU501215","term_id":"984874581","term_text":"KU501215"}}KU501215) was propagated in Vero cells (ATCC). The cells were grown in EMEM to 70% confluency in 25 mL tissue culture flasks (Nunc Inc., USA). Following removal of the culture media, virus inoculum was added to give a multiplicity of infection (MOI) of 0.1 to 0.05/cell or 1.0 to 1.5/cell. Flasks were incubated at 37 °C, 5% CO₂ with gentle agitation for 30 min. After incubation, 5 mL of complete media was added and the cells were maintained for 5–9 days or until cells exhibited cytopathic effects (CPE). Cells were trypsinized and cryopreserved in 50% FBS and 10% DMSO at − 80 °C and used in subsequent studies.