Fig 3. Lipin1 silencing interferes with early steps of HCV infection.

Panels A, B- Huh-7 cells were transduced with lentiviral vectors expressing control or LPIN1-specific shRNAs. At day 7 post-transduction, cells were infected at MOI 10 with HCV D183. Samples of the supernatants and the cells were collected at 24, 48 and 72 hours post-infection. (A) Relative infectivity titers found in infected cell supernatants. (B) Intracellular HCV RNA determined at the indicated time points is shown as genome equivalents per microgram of total RNA GE/μg). Panels C, D-Persistently infected cultures were generated by inoculation with JFH-1 virus at MOI 0.01. Once cultures reached >95% of HCV-positive cells, they were transduced with lentiviral vectors expressing control, HCV RNA-targeting or LPIN1-specific shRNAs. At day 7 post-transduction, cells were split and samples of the cells and supernatants were collected 24 hours later to determine infectious virus production rate by infectivity titration HCV (C) and RNA levels by RT-qPCR (D). All data are shown as mean and SD of 3 independent experiments performed in triplicate (n = 9). Statistical significance was determined using Student´s t-test (*p<0.05; **p<0.01).