Vero cells preseeded on 20-mm glass coverslips were infected with/without the NR-WT or NR-TC27 ZIKV at an MOI of 0.1. At the indicated time points, the culture supernatants were removed, and the cells were rinsed once in Opti-MEM medium (Invitrogen). For FlAsH/ReAsH (Invitrogen) labeling, the cells were incubated in 500 μL of labeling medium containing 2.5 μM FlAsH/ReAsH for 10 min at 37°C and were washed three times in Opti-MEM medium containing 5 × BAL (British anti-Lewisite) (each wash for 5 min). Subsequently, the cells were stained with Hoechst 33258 or fixed for IFA.
For FlAsH/ReAsH pulse-chase labeling, the infected cells were cultured in Opti-MEM containing 0.4 μM FlAsH/ReAsH. For live-cell imaging, the cell nuclei were stained using Hoechst 33258 for 10 min at 37°C and washed three times in Opti-MEM medium.
For FlAsH/ReAsH pulse-chase labeling, the infected cells were cultured in Opti-MEM containing 0.4 μM FlAsH/ReAsH. For live-cell imaging, the cell nuclei were stained using Hoechst 33258 for 10 min at 37°C and washed three times in Opti-MEM medium.