Infection of mouse fetal brain

Specific-pathogen-free ICR mice were purchased from Beijing Vital River Laboratory Animal Technology (licensed by Charles River). 6-week-old ICR mice were mated at embryonic day 0 (E0), and pregnant mice were selected for the ZIKV injection (1 μl of 1 × 10⁷ PFU/ml) into the cerebroventricular space of fetal brains at E13.5. The fetuses were inspected at E18.5 for the body weight, brain size and weight, and either homogenized and lysed for protein detection by immunoblotting or perfused with ice-cold 4% paraformaldehyde (PFA) for protein detection by Immunofluorescence assay (IFA) as described below. All experimental procedures involved were performed according to protocols approved by the Institutional Animal Care and Use Committee of Wuhan institute of Virology, Chinese Academy of Sciences.