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. 2003 May;77(9):5493–5498. doi: 10.1128/JVI.77.9.5493-5498.2003

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FIG.1. — Effects of IL-1 on HCV RNA replication and viral protein expression. (a) Northern blot analysis of total cellular RNAs from FCA1 cells treated with IL-1β. GAPDH served as an internal control. RNA from parental Huh-7 cells served as a negative control. The positive control was subgenomic HCV replicon RNA generated by in vitro transcription. (b) Graphic representation of the Northern blot results. Cells were treated with 0, 5, 10, and 20 ng of IL-1β per ml (from left to right for each set of bars). The HCV RNA levels from the control cells were set at 100% at each time point. The data were normalized to the internal control (GAPDH) values. (c) Immunohistochemical staining of cells treated with IL-1β. Cells were cultured on coverslips in the presence or absence of 10 ng of IL-1β per ml. Cells were fixed with 95% ethanol-5% acetic acid and stained with a monoclonal antibody against NS5A. The parental Huh-7 cells are shown for a negative control. Bar, 20 μm. (d) Western blot analysis. Approximately 50 μg of total protein from FCA1 cells treated with IL-1β or IFN-α for 24 h was resolved on a gel by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a membrane. FCA1 cells were treated with 10 (lane 3) or 20 (lane 4) ng of IL-1β per ml or with 10 (lane 5) or 50 (lane 6) U of IFN-α per ml. Huh7 (lane 1) and FCA1 (lane 2) cells not treated with IL-1β or IFN-α were controls. An anti-HCV NS5A monoclonal antibody was used to detect NS5A protein. Actin served as an internal control.

FIG.1. — Effects of IL-1 on HCV RNA replication and viral protein expression. (a) Northern blot analysis of total cellular RNAs from FCA1 cells treated with IL-1β. GAPDH served as an internal control. RNA from parental Huh-7 cells served as a negative control. The positive control was subgenomic HCV replicon RNA generated by in vitro transcription. (b) Graphic representation of the Northern blot results. Cells were treated with 0, 5, 10, and 20 ng of IL-1β per ml (from left to right for each set of bars). The HCV RNA levels from the control cells were set at 100% at each time point. The data were normalized to the internal control (GAPDH) values. (c) Immunohistochemical staining of cells treated with IL-1β. Cells were cultured on coverslips in the presence or absence of 10 ng of IL-1β per ml. Cells were fixed with 95% ethanol-5% acetic acid and stained with a monoclonal antibody against NS5A. The parental Huh-7 cells are shown for a negative control. Bar, 20 μm. (d) Western blot analysis. Approximately 50 μg of total protein from FCA1 cells treated with IL-1β or IFN-α for 24 h was resolved on a gel by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a membrane. FCA1 cells were treated with 10 (lane 3) or 20 (lane 4) ng of IL-1β per ml or with 10 (lane 5) or 50 (lane 6) U of IFN-α per ml. Huh7 (lane 1) and FCA1 (lane 2) cells not treated with IL-1β or IFN-α were controls. An anti-HCV NS5A monoclonal antibody was used to detect NS5A protein. Actin served as an internal control.