Quantifying ZIKV Infection Kinetics

Cells seeded at 5.7 × 10⁴ in 8-well chamber slide (Permanox; Nunc) were mock infected or infected with ZIKV at an MOI of 1. Immunocytochemistry was performed as previously described using 1:200 anti-flavivirus group E antigen antibody (clone 4G2; Millipore) or an isotypic control and 1:500 HRP-conjugated anti-mouse immunoglobulin secondary antibody (Cell Signaling Technology) (68 (link)). Stained cells representing infected cells were counted. The percent infection in live cells was calculated as the number of stained cells over total number of cells counted. Percent infection in live and dead cells was calculated as [(% viability at 24 h − % viability at 48/72 h) × 100 + (% viability at 48/72 h × % of infection in live cells at 48/72 h)]/% viability at 24 h.