Figure 2.

HCV transcriptionally suppresses miR‐30e promoter activity. (A) Huh7.5 cells (control) and Huh7.5 cells harboring the HCV genome‐length replicon (HCV replicon) were transfected with the miR‐30e‐luciferase reporter construct (P₀), and promoter activity was measured by a relative luciferase assay after 48 hours of transfection. Data are presented as mean ± SD from at least three independent experiments. (B) Schematic diagram of the miR‐30e promoter region (nucleotides −1813 to +1; P0) cloned into the luciferase reporter plasmid (P0) and a deletion mutant of the miR‐30e promoter construct (nucleotides −1567 to +1 [P1]). (C) Huh7.5 cells transfected with the miR‐30e‐luciferase reporter constructs (P₀ or P₁) knocked down C/EBP‐β using a specific siRNA, and promoter activity was measured by a relative luciferase assay after 48 hours of transfection. Data are presented as mean ± SD from three independent experiments. (D) Huh7.5 cells were left uninfected (mock) or infected with HCV for 48 hours. Cell lysates were analyzed for C/EBP‐β expression by western blot using specific antibodies. The blot was reprobed with antibody to actin for protein load. Densitometric scanning results are presented. (E) Huh7.5 cells transfected to control or siRNA to C/EBP‐β. Cell lysates were prepared 48 hours after transfection and analyzed for C/EBP‐β expression by western blot using specific antibodies. The blot was reprobed with antibody to actin for protein load. Densitometric scanning results are presented. Statistical significance was analyzed using the two‐tailed Student t test; *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: F luc, firefly luciferase; NS, not significant.