Figure 1.

Ligand-based stimulation of retinoic acid inducible gene I (RIG-I) by expression of Hepatitis C virus (HCV) RdRP NS5B. NS5B (a) and IFIT1 (b) mRNA levels in NS5B^WT, NS5B^mut, or empty vector transduced A549 cells, 6 days post transduction. NS5B fold change (f.c.) over GAPDH or IFIT1 fold change over GAPDH and empty vector control was calculated (mean + s.d. of n = 3 technical replicates). One representative of n = 3 independent experiments is shown. (c) IFN-λ mRNA levels in naïve cells or cells transduced with NS5B^WT or NS5B^mut either correspond to the entire cell pool or FACS sub-populations (P1-6), which were sorted by increasing NS5B^WT-eGFP levels. Fold change over GAPDH and naïve cells was calculated (mean + s.d. of n = 3 technical replicates). Dashed lines indicate IFIT1 mRNA levels of the cell pool for naïve or NS5B^WT transduced cells. (d) Total proteome analyses of A549 cells 12 days post NS5B^WT or NS5B^mut transduction. Hyperbolic lines indicate significantly changing proteins (two-tailed Student’s t-test, S0 = 1, permutation-based FDR < 0.01, n = 4 technical replicates). Proteins annotated with the GOBP term “cellular response to type I interferon” are highlighted in blue. (e) Bar plot showing Benjamini–Hochberg-adjusted (B.H.) FDR values (Fisher’s exact test) for the seven most significantly enriched GOBP terms within significantly changing proteins upon NS5B^WT expression. (f) Western blot of NS5B and IFIT1 protein expression in wild-type and RIG-I knock-out A549 cells transduced with NS5B^WT for the indicated days. Calnexin (CANX) was used as loading control.