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. 2019 Jul 3;7(3):59. doi: 10.3390/vaccines7030059

Figure 2.

Figure 2

Both, wild type and codon-optimized RSV F, are efficiently expressed in VSV-GP-F infected cells and RSV F is efficiently incorporated in VSV-GP-F particles. (A) BHK-21 cells were infected with an multiplicity of infection (MOI) of 0.1 or 0.01 with replication competent VSV-GP variants containing a wild type (VSV-GP-Fwt) or a codon optimized version of RSV F (VSV-GP-Fsyn). As negative control, cells were infected with VSV-GP without additional transgene. Then, 24 h after infection, cell lysates were prepared and analyzed by Western blot using RSV F-specific (top row), LCMV GP-specific (2nd row), VSV N-specific (3rd row), or actin-specific (lower row) antibodies; (B) virus stocks were produced on BHK-21 cells, concentrated via low speed centrifugation through a sucrose cushion and purified virus stocks of VSV-GP-Fwt and VSV-GP-Fsyn were analyzed via Western blotting using RSV F-specific (top row), LCMV GP-specific (middle row), or VSV N-specific (lower row) antibodies. As negative control, VSV-GP without additional transgene was used.