Tissues were fixed in 4 % paraformaldehyde for 1 h, then in 30 % sucrose solution overnight for cryoprotection. 10–12 μm thick sections were cut using a Leica cryostat. Immunohistological detection of HBcAg was performed on frozen sections using primary rabbit polyclonal anti-HBcAg antibody (1:200; Abcam, England) and Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200; Jackson ImmunoResearch, USA). Finally, the sections were stained with 4′,6-diamidino-2-phenylindole (DAPI) for nuclear indication. Immunofluorescence analyses were performed with a fluorescence microscope (Leica, German).
HBV core protein was also analyzed by a standard western blot procedure using primary polyclonal anti-HBcAg antibody (1:200; Abcam, England), anti-actin monoclonal antibody (1:1000; Santa Cruz Biotechnology, USA) and peroxidase-conjugated secondary antibodies. The image was digitized using a scanner and signal was quantified using of Quantity One software (Bio-Rad).
HBV core protein was also analyzed by a standard western blot procedure using primary polyclonal anti-HBcAg antibody (1:200; Abcam, England), anti-actin monoclonal antibody (1:1000; Santa Cruz Biotechnology, USA) and peroxidase-conjugated secondary antibodies. The image was digitized using a scanner and signal was quantified using of Quantity One software (Bio-Rad).