Effect of CP-346086 at low concentrations on apoB and apoE secretion and HCV production in multiple-cycle HCV growth assays. Effect of CP-346086 at low concentrations on apoB and apoE secretion and HCV production in multiple-cycle HCV growth assays. Huh7.5 cells were infected with HCV at an MOI of 0.3 and then incubated with DMEM containing CP-346086 at concentrations varying from 0.008 to 1 μM. After 3 days p.i., the medium was collected for determination of apoB and apoE secretion and HCV production. (A) Determination of apoB and apoE secretion by Western blotting. (B) Determination of infectious HCV by infectivity assay. HCV in the medium was used to infect naïve Huh7.5 cells. At 3 days p.i., the level of HCV NS3 protein was determined by Western blotting. (C) Correlation of apoB-100 and apoE secretion with HCV production. The levels of apoB-100, apoE, and NS3 in panels A and B were quantified and converted to percentages of the control levels, considering the levels of apoB-100, apoE, and NS3 in the absence of CP-346086 as 100%. Relative levels of apoB-100, apoE, and NS3 were plotted against concentrations of CP-346086. (D) Determination of infectious HCV titers by IFA. Infectious HCV in the medium of HCV-infected and inhibitor-treated cells was titrated and quantified by IFA as FFU/ml. Jieyun Jiang, and Guangxiang Luo J. Virol. 2009;83:12680-12691