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. 2018 May 18;42(2):957–965. doi: 10.3892/ijmm.2018.3693

Figure 3.

Figure 3

Attenuation of IFN-α signaling by type I collagen is β1-integrin-dependent. (A) β1-integrin was overexpressed in Huh7 cells grown on type I collagen-coated dishes. (B) Improvement in the ISRE luciferase activity after treatment with β1-integrin function-blocking antibody in HuH-7 cells cultured on type I collagen-coated dishes. Cells were treated with β1-integrin function-blocking antibody (1 µg/ml) for 6 h. The ISRE-luciferase activity was measured after IFN-α treatment for 12 h. The results are presented as the mean fold induction of the controls. (C) Improvement in the ISG protein expression by treatment with β1-integrin function-blocking antibody in HuH-7 cells grown on type I collagen-coated dishes. The cells were cultured on type I collagen-coated dishes for 3 days and then treated with β1-integrin function-blocking antibody for 6 h, followed by treatment with IFN-α for 12 h. The ISG15 and PKR expression was measured by western blot analysis with β-actin used as a control. (D) Improvement in the suppressive effect of IFN-α on HCV replication in OR6 cells cultured on type I collagen-coated dishes. After 6-h treatment with β1-integrin function-blocking antibody (1 µg/ml), the cells were treated with IFN-α for 12 h, and the Renilla luciferase activity was then measured. (E) Improvement in the suppressive effect of IFN-α on HCV protein expression. HCV-NS5a expression was suppressed by co-treatment with β1-integrin function-blocking antibody in OR6 cells grown on type I collagen-coated dishes. The OR6 cells were cultured on type I collagen-coated dishes for 3 days and then treated with β1-integrin function-blocking antibody for 6 h, followed by treatment with IFN-α for 12 h. The HCV-NS5a expression was measured by western blot analysis with β-actin used as a control. Values are expressed as the mean ± standard deviation (n=3). *P<0.05. IFN, interferon; OR6 cells, HuH-7 cells stably transfected with full-length HCV-RNA fused with Renilla luciferase HCV, hepatitis C virus; NS, nonstructural protein; ISRE, IFN-stimulated response element; ISG, IFN-stimulated gene; PKR, protein kinase R.