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Fig. 4 | Virology Journal

Fig. 4

From: Identification of a novel interaction site between the large hepatitis delta antigen and clathrin that regulates the assembly of genotype III hepatitis delta virus

Fig. 4

W207 situated in the C terminus of HDV-III HDAg-L enables binding to CHC. A Binding of His-CHC(1–107) to GST-III(198–210) or GST-III(198–210)-W207A. GST-tagged proteins were incubated with His-CHC(1–107). Levels of His-CHC(1–107) were determined by western blot analysis using anti-His antibody. Coomassie blue staining of GST fusion proteins is shown in the bottom panel. B Coimmunoprecipitation of CHC with HDV-III HDAg-L. The lysates of Huh7 cells cotransfected with plasmids pECE-C-ES and pCMV-Tag2C-HDAgS-III, pCMV-Tag2C-HDAgL-III, or pCMV-Tag2C-HDAgL-III-W207A were incubated with the anti-CHC antibody for precipitation. Levels of CHC, Flag-tagged HDAgs, and small HBsAg were determined by western blot analysis using anti-CHC, anti-Flag, and anti-HBsAg antibodies. (C) Secretion of HDV-III VLPs. HDV-III VLPs were harvested from the medium of Huh7 cells cotransfected with plasmids pECE-C-ES and pCMV-Tag2C-HDAgS-III, pCMV-Tag2C-HDAgL-III, or pCMV-Tag2C-HDAgL-III-W207A at day 4 posttransfection. Levels of Flag-tagged HDAgs and small HBsAg were determined by western blot analysis using anti-Flag and anti-HBsAg antibodies. Experiments were performed in triplicate

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