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. 2014 May 15;10(5):e1004128. doi: 10.1371/journal.ppat.1004128

Figure 4. HTEIs effectively block cell-cell transmission of DAA-resistant viruses.

Figure 4

The experimental setup is shown in Figure 1C. NS5A+ HCV producer cells (Pi) were transfected with HCV RNA encoding for HCV Jc1 NS3-A156S (A-B) or Jc1 NS3-L36M/R155K (C–D). NS5A+ HCV producer cells and GFP-expressing target cells (T) were co-cultivated with nAb (anti-HCV IgG, 25 µg/mL) to block cell-free transmission as described [6]. Cell-cell transmission of wild-type or drug-resistant strains was determined by quantification of GFP+ NS5A+ target cells (Ti) by flow cytometry. Protease or NS5A inhibitor-resistant HCV variant producer cells (Pi) cultured with uninfected target cells (T) were then incubated with 1 µg/mL of CLDN1-specific mAb or 10 µM of erlotinib or control medium. HCV-infected target cells (GFP+NS5A+) were quantified by flow cytometry (A and C). Percentage of infected target cells is shown as histograms (B and D) and is represented as means ± SD from three experiments performed in triplicate. *p<0.005.