Fig 5. Lipin1-silencing interferes with early HCV RNA replication, downstream of viral entry and primary translation.
Huh-7 cells were transduced with lentiviral vectors expressing control or LPIN1-specific shRNAs. (A) Control and silenced cells were inoculated with HCV E1E2 (HCVpp) or VSV-G-pseudotyped retroviral vectors (VSVpp) 7 days post-transduction. Control cells treated with hydroxyzine (5μM) were used as positive inhibition control. Luciferase activity was determined in the different cell lines at 48 hours post-inoculation. HDX, hydroxyzine pamoate (5μM). Panels B, C- Control and lipin1-deficient cells were transfected with a replication-deficient mutant (B) or replication competent subgenomic HCV replicon bearing a luciferase gene (C). Luciferase activity was determined in the different cell lines at 5 hours post-transfection for both replicons and 48 hours post-transfection for the replication-competent replicon RNA. Data are expressed as average and SD of three independent experiments performed in triplicate (n = 9). Statistical significance was determined using Student´s t-test (*p<0.05; **p<0.01).