Figure 6. A Recombinant RVFV Which Does Not Interact with SAP30 Induces IFN-β and Is Avirulent.

(A) Monolayers of Vero cells infected with RVFV rec-ZH or rec-ZHΔ210–230 were fixed and stained with crystal violet at 5 days p.i. (B) GST or GST-SAP30 was incubated with extracts from cells uninfected or infected with rec-ZH or rec-ZHΔ210–230. Proteins from the crude extracts (input) or after binding on GST or GST-NSs beads were analyzed by Western blotting using anti NSs antibodies. * denotes a cellular protein bound on GST-NSs which copurified with NSs. BF (C) or L929 wt 330 cells (D, E, F) uninfected or infected with C13, wt ZH, rec-ZH, or Δ210–230 were incubated for 8 h (C), 18 h (E) or for the indicated time (D and F). Extracts were prepared and analyzed by RT-PCR to detect C) IFN-β, GAPDH, or NSs mRNA as described in [26]. (D) CAT activity, (E) confocal microscopy, or (F) chip experiment using anti-NSs antibodies like in Figure 3; (G) percentage of animals surviving after i.p. inoculation of 10⁴ pfu.