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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 7

Effect of RVFV NSs on host cell factors.

A. Effect on PKR and p62. A549 cells or A549 cells treated with 5 µg/ml actinomycin D were infected with rMP12 or rMP12:S-Swap at a MOI of 3, or mock-infected. Cell extracts were prepared at the time points indicated, proteins fractionated by SDS-PAGE, and blots probed with anti-N, anti-NSs, anti-PKR, anti-p62, and anti-tubulin antibodies as indicated. B. Induction of interferon. A549 cells were infected with rMP12, rMP12ΔNSs:eGFP, rMP12:S-Swap or rMP12:S-SwapΔNSs:eGFP at MOI of 5, and supernatants harvested at 18 h p.i. After UV treatment, 2-fold dilutions were applied to A549-NPro cells for 24 h, before infection with EMCV. Monolayers were stained with Giesma after a further 96 h. C. The amount of IFN produced is expressed as relative IFN units (RIU), defined as RIU = 2N where N = the number of two-fold dilutions of the supernatants that protected the reporter cells from EMCV challenge.

Figure 7