Characterization of anti-HCV activities of N-89 and N-251.

<p>(A) Schematic gene organization of authentic HCV-RNA (HCV-O/RLGE). The positions of four adaptive mutations - Q1112R, P1115L, E1202G, and K1609E - are indicated by a black star. (B) N-89 and N-251 inhibited authentic HCV-RNA replication. The cells harboring HCV-O/RLGE RNA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072519#pone.0072519-Ikeda4" target="_blank">[19]</a> were treated with N-89 (left panel) and N-251 (right panel) for 72 hrs, followed by real-time LightCycler PCR (black circles in the upper panel) and WST-1 assay (open triangles in the upper panel). The relative value (%) calculated at each point, when the level in non-treated cells was assigned as 100%, is presented here. Data are expressed as the means±standard deviation of triplicate assays. Western blot analysis (lower panels) was performed as described in Fig. 1B. (C) N-89 did not inhibit the HCV-JFH-1 replication. RSc (left panel) and D7 (right panel) cells were inoculated with supernatant from RSc cells replicating JR/C5B/BX-2 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072519#pone.0072519-Takeda1" target="_blank">[29]</a>. The RL assay was performed as described in Fig. 1B. (D) N-89 (left panel) and N-251 (right panel) did not inhibit the replication of HCV-JFH-1 subgenomic replicon. The RL and WST-1 assays were performed as described in Fig. 1B.