Immunohistochemical analysis
For immunohistochemical analysis, tissue samples (including spleens,
tonsils, and gastrohepatic lymph nodes) were collected from ASFV
infected pigs and fixed in 10% formalin for 24 h at room temperature.
Slides were prepared following a common procedure of dehydration,
embedding, and section. The slides were dried in oven at a temperature
of 56℃ for 15 min, incubated with 3% perhydrol liquid at room
temperature and blocked with 5% Difco Skim Milk for 1h at room
temperature, incubated with 1:100 of mAbs at 4℃ overnight. After five
washes with TBST, the slides were incubated with a goat anti-mouse IgG
(Invitrogen, USA) (1:100) for 30 min
at 37℃. The slides were stained with ST Hematoxylin (Leica, Germany) and
analyzed using a microscope (DFC550, Leica, Germany).