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. 2011 Jul 21;7(7):e1002144. doi: 10.1371/journal.ppat.1002144

Figure 4. Plasmid constructs and expression levels of core and E2 proteins.

Figure 4

Schematic diagrams of HCV proteinexpression constructs used in this study A. The sequences encoding core to the first TMS of NS2 derived from the J6CF HCV isolate are indicated in gray boxes whereas sequences from JFH1 strain are shown in white boxes. Core, coreE1E2 CE2, coreE1E2p7 Cp7, coreE1E2p7NS2 CNS2 polyproteins derived from JFH1 and J6CF Jc1 isolates were expressed using a CMV promoter expression construct. The p7 and p7NS2 proteins were expressed using a signal peptide derived from the last 45 aminoacids of HCV E2 E2 via transduction with MLVbased retroviral vectors. At 72h posttransfection, lysates from mocktransfected cells, from JFH1 or Jc1 HCVccexpressing cells, from Huh7.5 cells transfected with the JFH1 or Jc1 core, CE2, Cp7 or CNS2 expression constructs, or from Huh7.5 cells transduced with the MLVbased vectors expressing JFH1 or Jc1 p7 or p7NS2, as indicated, were prepared and examined by Western blot analysis using antibodies against NS2, core and E2 proteins B. The input of the samples was assessed by staining with an Actin antibody.