Following a virus cross-neutralization test against FAdV-8a and -8b reference strains, as described above, the cells were fixed by adding ice-cold methanol for 5 min. Following serial washing steps and blocking with 3% bovine serum albumin (BSA) for 1 h, in-house-generated polyclonal rabbit anti-FAdV antiserum (diluted 1:500 in PBS) was added to each well overnight. After removal of the rabbit antiserum and washing, the plates were incubated with 1:200-diluted Alexa Fluor 488-conjugated donkey anti-rabbit IgG (Invitrogen, Life Technologies, Carlsbad, CA, USA) in the dark for 1 h. After another wash, cells were stained with 1:1,000 DAPI (4′6-diamidino-2-phenylindole solution; Roche Diagnostics GmbH, Vienna, Austria) for 5 min, subjected to a final wash, and covered with PBS.
Internalization of viral particles was examined and documented with a Zeiss Axiovert 200 M fluorescence microscope (Zeiss, Jena, Germany) coupled to a Flexacam C1 camera and Leica Application Suite X (LAS X) (Leica Microsystems GmbH, Wetzlar, Germany).
Internalization of viral particles was examined and documented with a Zeiss Axiovert 200 M fluorescence microscope (Zeiss, Jena, Germany) coupled to a Flexacam C1 camera and Leica Application Suite X (LAS X) (Leica Microsystems GmbH, Wetzlar, Germany).