Figure 1.

Upregulation of HULC expression by hepatitis C virus (HCV) replication in cell culture. A, Huh-7.5 cells were infected with cell culture-derived HCV of HJ3-5 at multiplicities of infection (MOIs) of 0.01, 0.1, and 1 or mock. Total RNA was extracted at 24, 48, 72, and 96 hours postinfection. HCV RNA, HULC, and 18S ribosomal RNA (rRNA) levels were quantified by reverse-transcription quantitative polymerase chain reaction (RT-qPCR), and the levels of HCV RNA and HULC were normalized to those of 18S rRNA, and further normalized to the relative HULC level of mock at 24 hours, which was set to 1. Error bars show the standard deviation from 3 independent experiments, and the differences of means among each condition were analyzed by 2-way analysis of variance (ANOVA). B, Schematic representation of full-genomic and subgenomic replicons. Neo-R, neomycin resistance gene. C, Huh-7.5 cells harboring each replicon were established after selection with 0.5 mg/mL G418 for 21 days, and total RNA was extracted. The levels of HCV RNA, HULC, and β-actin were quantified by RT-qPCR, and HCV RNA and HULC levels were normalized to those of β-actin. Differences in the means were analyzed by one-way ANOVA. D, Huh-7.5 cells harboring the M1LE replicon were treated with 5 μM sofosbuvir or 0.5% dimethyl sulfoxide (DMSO) without neomycin. HCV RNA, HULC, and 18S rRNA levels were determined by RT-qPCR every other day, and HCV RNA and HULC levels were normalized to the level of 18S rRNA. *P < .05, **P < .01, ***P < .001.