Preparation of DEN2-NGC and YFV-17D stocks has been previously described [8] (link). Infectious stocks of JFH1 HCV were generated and titrated by foci forming assay using HuH-7.5 cells as described in [16] (link).
Titers of stocks and experimental samples were determined by foci forming assays. These assays were performed as described in Sessions et al. [8] (link) with the following modifications: 2×105 Vero cells/well were plated in 24-well plates. Following virus adsorption, 0.5 mL of 1∶1 1.2% Tragacanth Gum (Sigma)/2× EMEM (Lonza), supplemented with 5% FBS and 10 mM HEPES (Invitrogen) was added per well. The primary antibodies 4G2 and YF-mAF were used to detect DEN2-NGC and YFV-17D, respectively. Alexa488-conjugated anti-mouse antibody was used as secondary and foci were detected by immuno-fluorescence.
Titers of stocks and experimental samples were determined by foci forming assays. These assays were performed as described in Sessions et al. [8] (link) with the following modifications: 2×105 Vero cells/well were plated in 24-well plates. Following virus adsorption, 0.5 mL of 1∶1 1.2% Tragacanth Gum (Sigma)/2× EMEM (Lonza), supplemented with 5% FBS and 10 mM HEPES (Invitrogen) was added per well. The primary antibodies 4G2 and YF-mAF were used to detect DEN2-NGC and YFV-17D, respectively. Alexa488-conjugated anti-mouse antibody was used as secondary and foci were detected by immuno-fluorescence.
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