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. 2009 May 15;284(20):13348–13354. doi: 10.1074/jbc.M809449200

FIGURE 3.

FIGURE 3.

Characterization of RIG-I S183I. A, RIG-I–/– MEF cells were transfected with reporter genes together with empty vector (Vector) or plasmid expressing FLAG-tagged WT RIG-I, RIG-I CARD (the N-terminal region, amino acid 1–229), or RIG-I CARD S183I. After transfection (24 h), the cells were subjected to a Dual-Luciferase assay. Error bars show the S.D. values for triplicate transfections. B, each protein expressed in RIG-I–/– MEF cells was detected by immunoblotting using an anti-FLAG antibody. RIG-I full, full-length RIG-I. C, reporter assay of the K172R mutant was performed as in Fig. 2B. D, protein levels were determined by immunoblotting. RIG-I K172R and WT RIG-I were expressed at comparable levels. E, empty vector or expression vectors for full-length RIG-I with the T55I or S183I mutation were introduced into L929 cells (the total amount of plasmid was kept at 9 μg by adding empty vector) and infected with Newcastle disease virus, and reporter activity was analyzed as in panel A. To observe the dose response, cells received 1, 5, or 9 μg of the expression plasmid for T55I and S183I as indicated. NDV, Newcastle disease virus.