2.4 Western blot analysis
Cells (HEK293T) were transfected with the two-plasmid system and incubated at 37 °C until the pseudotyped EBOV was harvested. The cells were harvested and lysed. After centrifugation, the lysates were mixed with 4×LDS sample buffer (ThermoFisher Scientific) and denaturing at 90℃for 10 min. Samples were loaded onto to a 12% SDS-PAGE gel. After electrophoresis, proteins were transferred onto nitrocellulose (NC) membranes. The NC membranes were then blocked using SuperBlocking Buffer (Thermo Fisher, TX, USA) at room temperature for 2 h and subsequently incubated with anti-p24 antibody (Sino Biological, China) and anti-EBOV GP antibody (rabbit serum immunized with purified, truncated EBOV GP inEscherichia coli ) for 1 h at room temperature. After three washes with phosphate-buffered saline containing tween 20 (PBST), the NC membranes were incubated with HRP-conjugated goat anti-rabbit IgG (Bios, China) at room temperature for 1 h. The signals were visualized on a Fujiflm LAS-4000 image reader (Fujiflm, Tokyo, Japan) with SuperSignal West Dura Extended Duration Substrate Kit (Thermo Fisher).