FIGURE 1.

Impaired induction of ARE-regulated genes in HCV-replicating cells. A, reporter gene assay in Huh7.5 cells electroporated with HCV replicons JFH1wt or JFH1/J6. Cells electroporated with the replication-deficient construct JFH1_GND served as control. 48 h after electroporation cells were co-transfected with the reporter constructs harboring the NQO1, GI-GPx, or GCLM-derived ARE sequences. Activities are indicated as mean values from three independent experiments. The bars represent the standard deviation. Cells were stimulated with tert-butylhydroquinone (tBHQ) (black bars) or left unstimulated (gray bars). *, p < 0.05). B, analysis of NQO1-expression by reverse transcription polymerase chain reaction (RT-PCR). Electroporated cells were incubated with 60 μm tBHQ for 16 h or treated with the equivalent amount of DMSO as control. Amplification of HCV- and actin-specific sequences served as control. C, Western blot analysis of cellular lysates derived from HuH7.5 cells electroporated with HCV replicons JFH1wt or JFH1/J6 using a NQO1-specific antiserum. Cells electroporated with the replication-deficient construct JFH1_GND served as control. 96 h after electroporation cells were stimulated for 16 h with 60 μm tBHQ (right panel). HCV replication was visualized by a NS5A-specific serum. D, analysis of GPx and GCLC expression normalized to GAPDH in HCV-infected primary human hepatocytes. Uninfected PHH served as control. Expression was analyzed by real time PCR. The data represent mean values from three independent experiments and are shown as relative values. The bars represent the standard deviation. E, analysis of GPx- and GCLC expression normalized to GAPDH in liver samples derived from three patients with chronic hepatitis. Liver samples from three HBV-/HCV-negative patients served as control. Expression was analyzed by real time PCR. The data represent mean values and are shown as relative values. The bars represent the standard deviation.