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. 2007 Sep 5;81(22):12227–12237. doi: 10.1128/JVI.01300-07

FIG. 5.

FIG. 5.

pppRNA-induced activation of IFN-β. (A) Parallel cultures of MEFs were transfected with pIFNβ-lucff and pTK-lucr, and pRIG-I was also transfected in some cultures, as indicated. After 24 h, the cells were transfected for 3 h with increasing amounts (1 or 3 μg) of either pppGGG/RNA1, phosphatase-treated GGG/RNA1, pppGCA/RNA1, or OHGCA/RNA1, as indicated. Cytoplasmic extracts were prepared 18 h post-RNA transfection and used to determine luciferase levels. (B) Parallel cultures of MEFs were transfected with pIFNβ-lucff, pTK-lucr, and plasmids expressing Tom, RIG-I(ΔCARDs), IAV NS1(1-73), C1-23-Tom-C24-204 (C*), or V, as indicated. After 24 h, the cells were transfected with increasing amounts (1 μg and 3 μg) of pppGGG/RNA1, as indicated. Cytoplasmic extracts were prepared after 3 h of RNA transfection and used to determine luciferase levels. Rel., relative. (C) Same as above (B), except that the cells were transfected with 3 μg of ppptrailer RNA.