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The Consequences of Reconfiguring the Ambisense S Genome Segment of Rift Valley Fever Virus on Viral Replication in Mammalian and Mosquito Cells and for Genome Packaging

Figure 4

Intracellular localization of NSs in rMP12- or rMP12:S-Swap-infected cells.

A. Vero-E6 cells were infected at MOI of 1, and at the time points indicated the cells were fixed with 4% paraformaldehyde, followed by co-staining with anti-NSs (green) and anti-tubulin (red) antibodies. Cells were examined with a Zeiss LSM confocal microscope. B. Width of NSs filaments. The widths of NSs filaments (50) in rMP12- or rMP12:S-Swap were measured using the LSM Image Browser Software (Carl Zeiss MicroImaging GmbH) and the results presented as the mean width of the filaments and SEM of the two groups (**** = p<0.0001; see Methods). C. Detection of NSs in mosquito cells infected with rMP12. A.albopictus C6/36, U4.4 or A. aegypti Ae cells were infected with recombinant viruses (MOI of 1), and at 48 h p.i. the cells were fixed with 4% paraformaldehyde, followed by co-staining with anti-NSs (green) and anti-tubulin (red) antibodies (upper panels). Duplicate monolayers were stained with anti-N antibodies (lower panels). D. Detection of NSs in mosquito cells infected with rMP12:S-Swap. Cells were infected and stained with anti-NSs (green) and anti-tubulin (red) antibodies as above.

Figure 4

doi: https://doi.org/10.1371/journal.ppat.1003922.g004