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. 2004 Feb 2;101(6):1743–1747. doi: 10.1073/pnas.0308340100

Fig. 3.

Fig. 3.

Antiviral effect requires new transcription and translation. (A) Differentiated HBV-Met cells were pretreated for 1 h with 10 μg/ml α-amanitin, followed by the addition of 500 units/ml IFN-β. Southern blot (SB) and Northern blot (NB) analyses of HBV DNA-replicative intermediates and 2′5′-OAS/GAPDH expression were performed at 12 h post-IFN-β treatment. α-Amanitin blocks both IFN-induced gene expression and the antiviral effect. (B) Differentiated HBV-Met cells either were pretreated for 1 h with 100 μg/ml cycloheximide, followed by the addition of 500 units/ml IFN-β [cycloheximide (CHX) (1)], or were pretreated with 500 units/ml IFN-β for 3 h before the addition of 100 μg/ml CHX (3). SB and NB analyses of HBV DNA-replicative intermediates and 2′5′-OAS/GAPDH expression were performed at 12 h post-IFN-β treatment. When added 1 h before IFN, cycloheximide inhibits the antiviral effect and reduces IFN-induced gene expression. When added 3 h after IFN, CHX blocks the antiviral effect without altering IFN-induced gene expression. Tg, transgene; RC and SS, relaxed circle and single-stranded DNA-replicative intermediate forms, respectively.