Fig 9.

The requirement of TIA-1, TIAR, and G3BP1 proteins in HCV infection is independent of their presence in stress granules. (A) Huh-7 cells were transduced with one or two lentiviruses as indicated, and 5 days later, the expression of target proteins was determined by Western blotting. (B) After downregulation was confirmed, the cells were infected with JFH-1 D183 virus at a high multiplicity of infection (MOI = 5), and the intracellular HCV RNA and infectivity titers and extracellular infectivity titers were determined 24 h later as indicated in Materials and Methods. The results shown are averages and standard deviations (mean and SD; n = 3) (C) Cells infected in parallel were fixed 72 h after inoculation, and the number of HCV-infected cells containing stress granules was determined by immunofluorescence and counting as described for Fig. 2B. The results displayed were confirmed in 2 independent experiments, each one performed in triplicate. The shRNA sequences used in these experiments were shTIA-1 no. 1, shTIAR no. 1, shG3BP1 no. 1, shPKR, and shCtrl.