Fig 7. Western blot analysis of purified vaccine and wtRV virions and viral proteins in the infected HUVEC.

Vero and HUVEC were infected with RA27/3 and RV-Dz at MOI = 5. Virions were pelleted from the culture media by ultracentrifugation and resuspended in an SDS-sample buffer. The cell monolayers were washed with PBS and proteins were then extracted with RIPA buffer. Proteins were separated by a 4–12% NuPage gel, either nonreducing (E1, C, β-actin) or reducing (E2), and then the blots were probed with rubella E1, E2 and C-specific MAb to identify RV structural proteins. The blots were also probed with β-actin MAb to demonstrate equal protein loading for the analysis of the cell lysates and show purity of the pelleted virions. Representative results of two independent experiments are shown.