Figure 6.

mTOR-IRF3 pathway triggers antiviral activity

(A) HEK293T cells were transfected with or without IRF3 (WT and S386A) as indicated followed by amino acids (AA) or fetal bovine serum (FBS) treatment. RT-PCR was performed to analyze IFNβ mRNA expression level. Data are represented as mean ± SD; ∗∗∗, p < 0.001, one-way ANOVA.

(B) In HEK293T cells, IRF3 was cotransfected with mTOR alone or combined as indicated. ISG54 and ISG56 mRNA expression were analyzed with RT-PCR. Data are represented as mean ± SD.

(C) In FKBP12−/− mouse fibroblasts, FKBP12-K45R, -K45Q, -K48R, and -K48Q variants were overexpressed followed by VSV infection. RT-PCR was performed to analyze VSV expression. Data are represented as mean ± SD.

(D) In mouse fibroblasts with SEV or no virus infection, anti-FKBP12-aK53 immunostaining and DAPI staining were visualized with confocal fluorescent microscope. Scale bar: 20 μm.

(E) Western blot analysis revealed that VSV protein product was only detected in WT but not in FKBP12−/− cells.

(F) WT or FKBP12−WT mouse fibroblasts infected with VSV, the infected cells were analyzed by flow cytometry.

(G) Mouse fibroblasts were infected with VSV-GFP followed by AGK2 (20 μM) treatment and the cells were analyzed with flow cytometry.