Western blot (WB) analysis
The recombinant baculovirus was inoculated into Sf9 cells at an MOI of
0.5, and the mixture was harvested on the 4th day. The
precipitate and supernatant were separated by centrifugation, and the
precipitate was sonicated to obtain sonicated supernatant. Samples were
denatured in loading buffer at 100°C for 5 min and separated by 10%
sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
Then, the separated proteins were transferred onto nitrocellulose (NC)
membranes (GE Healthcare, Little Chalfont, Buckinghamshire, UK) for
immunoblot analysis. A polyclonal rabbit anti-ZIKV-E antibody (1:700)
was used as the primary antibody, and HRP-conjugated goat anti-rabbit
IgG (1:5000) was used as the secondary antibody. SuperSignal West Dual
Chemiluminescent substrate (Pierce, Rockford, IL, USA) was added as the
chromogen, and bands were imaged using a Tanon-5200 Chemiluminescent
Imaging System (Tanon, Shanghai, China).