Inhibition of HCV infection by aptamers against NS2.

<p>(<b>A/B</b>) Effect of NS2-specific aptamers on viral RNA replication in HCV full-length replicon cell (A) or HCV-infected Huh7.5 cells (B). FL-neo (A) or HCV-infected Huh7.5 cells (B) were treated with aptamer NS2-1, NS2-2 or NS2-3 for 72 hours. Intracellular HCV RNA was detected with real-time PCR and normalized with GAPDH. The data represented means of three different experiments. (<b>C</b>) The effect of NS2 aptamer on cell viability. HCV-infected Huh7.5 cells were treated by 100 nM or 200 nM aptamers for 72 hours. The effect of aptamer on cell viability was measured by MTS assay. The data were normalized with the control and represented means of three independent experiments performed in triplicate. (<b>D</b>) Titration of HCV particles produced in HCV-infected Huh7.5 cells with aptamer treatment by FFU assay. HCV-infected Huh7.5 cells were treated as described in part B. The cell culture supernatants and intracellular virus particles were harvested and titered by FFU assay. The infectivity titers in the supernatant or inside the cells are the average of three different experiments. (<b>E</b>) Effect of aptamers on H77-S viral RNA replication (upper) and the production of viral particles (lower) in infectious cell culture system. H77-S–infected Huh7.5 cells were treated with 100 nM of each aptamer or library for 72 hours. Intracellular viral RNA was quantified by real-time PCR and normalized with GAPDH. The data represented means of three independent experiments. The cell culture supernatants were harvested and titered by FFU assay. The infectivity titers in the supernatant are the average of three different experiments. (<b>F</b>) Effect of aptamers on JC1 viral RNA replication (upper) and the production of viral particles (lower). JC1 RNA was in vitro transcript from the plasmid pFL-JC1. JC1 RNA was transfected into Huh7.5 cells and JC1 virus was collected from the cells. JC1 virus was used to infect Huh7.5 cells in the presence of 100 nM aptamer for 72 hours. Intracellular viral RNA was quantified by real-time PCR and normalized with GAPDH. The data represented means of three independent experiments. The cell culture supernatants were harvested and titered by FFU assay. The infectivity titers in the supernatant are the average of three different experiments. (<b>G</b>) Effect of NS2-specific aptamers on hepatitis B viral DNA replication. HepG2.2.12 cells were inoculated with 100 nM of aptamer for 72 hours. Intracellular HBV DNA was detected with real-time PCR and normalized with GAPDH. The data represented means of three different experiments. ^*<i>P</i><0.05, ^**<i>P</i><0.01 verse library.