FIG 8.

Examining type III IFN signaling in Vero cells using Western blotting. Vero cells were infected with dolphin (T), bovine (B), or human (H) PIV-3 for 24 h. At 24 h postinfection, Vero cells were stimulated with recombinant IL-29/-28A/-28B (1 μg/ml) for 30 min to drive signaling downstream of the type III IFN receptor. (A) Western blotting was used to examine Stat1 and Stat2 phosphorylation during PIV-3 infection. β-Actin was used as a loading control. (B) Densitometry quantification from Western blotting in Vero cells was calculated. Relative quantification from band intensity was calculated by its expression relative to the level of the IFN-λ-stimulated positive control. This RQ was then normalized to the level of the appropriate total Stat1 or Stat2 to give the percent pStat1 to total Stat1 or percent pStat2 to total Stat2 value. A significant change in phosphorylation levels from those in the IFN-λ-treated positive control is indicated (*, P < 0.05).