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. 2013 Jan 10;9(1):e1003133. doi: 10.1371/journal.ppat.1003133

Figure 9. SBVSΔNSs induces the production of IFN.

Figure 9

A. IFN protection assay. 2fGTH cells were infected with the indicated viruses and supernatants were collected 24 h post infection. Supernatants were UV treated to remove infectious virus and fed to CPT-Tert cells after serial dilution. CPT-Tert cells were later infected with EMCV and the presence of CPE monitored and compared to cells supplemented with known amounts of universal IFN. B. The induction of IFN-β mRNA was investigated by RT-PCR from RNA extracted from 2fGTH cells infected with virus as indicated or transfected with Poly I∶C as a positive control. To control for the presence of residual genomic DNA all the samples were amplified after reverse transcription performed without reverse transcriptase (top panel, indicated as+/−RT). The quality of the extracted RNA was verified by the amplification of the 45S ribosomal RNA (middle panel). The presence of virus was confirmed by the amplification of part of the SBV S segment (bottom panel). C. IFN protection assay performed in primary ovine trophoblast cell (oTr-1) and primary ovine endothelial cells as described in A. D. Survival plots of 7 day old IFNAR(−/−) mice inoculated intracerebrally with sSBV, SBVSΔNSs or cell culture media as a control. Data indicate that SBVΔNSs is as virulent as sSBV in these mice that lack an intact IFN system.