Zika Virus Infection Dynamics in Neural Progenitor Cells

NPCs grown on coverslips coated with poly-D-lysine were infected with ZIKV at an MOI of 3 or 0.1, and mock-infected NPCs were used as a control. Coverslips were collected at the indicated time points post-infection and processed for IFA as described previously (Liu et al., 2017 (link)). For cryosections, perfused tissues were fixed in 4% PFA, dehydrated in 30% sucrose, and frozen in tissue freezing medium (Leica Biosystems, Germany). Coronal sections (thickness of 40 μm) were used for immunofluorescence staining as described previously (Tang et al., 2015 (link)). Mouse monoclonal antibodies for DCX (Abcam, Cat#ab18723), Tbr1 (Abcam, Cat#ab31940), Ctip2 (Abcam, Cat#18465) and anti-ZIKV human serum (Ma et al., 2017 (link)) were used as primary antibodies. Secondary antibodies included FITC-anti-mouse-IgG2a (Invitrogen, Cat#11-4210-82), Alexa Fluor 488 goat-anti-human-IgG (Invitrogen, Cat#A-11013) and TRITC-anti-mouse-IgG1 (Southern Biotechnology, Cat#1070-03). Nuclei were counterstained with DAPI (Life Technologies). Images were obtained using a two-photon microscopy with the UltraVIEW VoX 3D Live Cell Imaging System (Perkin Elmer).