Peroxidase substrate kit

Briefly, paraffin sections were dewaxed and rehydrated as described above. Antigen retrieval was performed by incubating sections in 10 mM citrate buffer (pH 6.0) in a decloaking chamber for 30 min at 95°C. Sections were blocked with 5% normal goat serum and then incubated with anti-PDCoV-N, MUC-2, Notch-1, or HES-1 (1:100) overnight at 4°C in a humidified chamber. For negative controls, sections were incubated in buffered saline with Tween 20 (PBST) only. An SABC-POD kit (rabbit or mouse IgG) and a peroxidase substrate kit (both from BOSTER, Wuhan, China) were used, respectively, for amplification and visualization of signal.

Antibodies PE-conjugated co-stimulatory molecules cluster of differentiation (CD) 80/86 (ab69778), PE-conjugated CD1a(ab25599), FITC-conjugated Workshop Cluster 3a (SWC3a, ab24885) were obtained from Abcam (New Territories, Hong Kong). FITC-conjugated histocompatibility leukocyte Ag II-DR (MHCII, LS-C129867) was obtained from LifeSpan BioSciences (Seattle, WA, USA). PE-conjugated SWC3a (74-22-15A) was obtained from BD Biosciences (USA). Recombinant swine interleukin 4 (IL-4), was bought from BioSource (USA). Recombinant swine granulocyte-macrophage colony-stimulating factor (GM-CSF), was bought from Invitrogen (USA). Lipopolysaccharide (LPS) was bought from Sigma (Japan). RPMI 1640 medium and 4,6-diamidino-2-phenylindole (DAPI) solution were bought from Invitrogen (Grand Island, NY, USA). Fetal bovine serum (FBS) was bought from MULTICell (Canada). SABC-POD (rabbit IgG) Kit and Peroxidase Substrate Kit were bought from BOSTER (Wuhan, China). RNA extraction kit was bought from TianGen Biotech (Beijing, China).