Fig. 2

Interaction of HDAg and Smad3. a Huh7 cells expressing L-HDAg or S-HDAg of three genotypes were IP’d with anti-Smad3 Ab, and the IP’d lysates were subjected to SDS-PAGE and immunoblotting with antiserum from delta antigen hepatitis patients or with anti-Smad3 Ab. Densitometric analysis were indicated. Results shown are mean ± SD from five independent experiments. *: p < 0.05; **p < 0.01. b Colocalization analysis of L-HDAg or S-HDAg (green) and the Smad3 (red) using double-immunofluorescence staining. Detection by monoclonal antibody against HDAg, both L-HDAg and S-HDAg (green) showed a high degree of colocalization with Smad3 (red), particularly in the nucleoli region. c L-HDAg or S-HDAg associations with Smad3 were investigated by in situ proximity ligation assay (PLA). Each PLA signal is visualized as a red fluorescent spot, and represents one detected association event between HDAg and Smad3. Incubation of HDAg antibody alone was a negative control. Cell nuclei were stained with DAPI (blue), corresponding phase contrast images are also shown