FIGURE 2.

Rapalog restores the antiviral activity of FRB*-MX1 in a dose-dependent way in the presence of F-NLS-FKBP. A, HEK293T cells were transfected with PB1, PB2, PA, and NP expression plasmids (25 ng each), pHW-NSLuc (100 ng), and pRL-CMV (25 ng). The cells were co-transfected with 150 ng of pCAXL, 75 ng of F-NLS-FKBP, 75 ng of FRB*-MX1, or 75 ng of F-NLS-FKBP and 75 ng of FRB*-MX1, as indicated. Six hours after transfection, different doses of rapalog were added to the cells. The normalized luciferase activity in the lysates was determined 24 h after transfection. Rel Luc activity, relative luciferase activity. The experiment was performed in technical triplicates, and the data are represented as mean ± S.E. The statistical probability that differences were significant between pairs of different constructs at a fixed rapalog concentration was assessed by a Fisher's protected least significant difference test. ****, p < 0.0001. B, representative Western blotting analysis of FRB*-MX1, NP, and F-NLS-FKBP in lysates of cell transfected as in A. The results shown in this figure are representative of two independent experiments.