To block non-specific binding, sections were treated with 4% BSA for 30 minutes.
To block non-specific protein binding, sections were treated with 3% normal goat serum and 0.05% Tween-20 (Biorad; Hercules, CA) in 1x APK buffer (Ventana Medical Systems, Inc.; Tucson, AZ) for 20 minutes at ambient temperature.
To block nonspecific binding, the sections were treated with 3% donkey serum in TBS with 0.1% Triton X-100 for 30 min (Jackson Immunoresearch Laboratories Inc, West Grove, PA, USA).
In order to prevent non-specific binding of CBM, sections were treated with a blocking solution containing 5% (w/v) ovalbumin in 50 mM sodium phosphate buffer (pH 7.4) for 1 h at rt followed by washing (twice, 15 minutes) in the same buffer.
To rule out non-specific binding of the lectins, tissue sections were treated, prior to lectin staining, with sialidase A (N-acetylneuraminate glycohydrolase; Prozyme) for 24 h at 37°C (pH 6.0), which preferentially cleaves all non-reducing terminal sialic acid residues in the order α(2,6) > α(2,3) > α(2,8) > α(2,9).
For staining ionized calcium binding adaptor molecule 1 (Iba-1), sections were treated with antigen retrieval reagent and then incubated with rabbit anti-Iba-1 antibody (Wako Pure Chemical Industries, Ltd., Osaka, Japan) overnight at 4°C.
After adequate antigen retrieval, the sections were treated for non-specific binding with 3% bovine serum albumin in PBS for 20 min and then incubated for 1 h at room temperature with primary antibodies.
To prevent the binding of nonspecific proteins to the primary antibody, the sections were treated with Protein Block Serum-Free (Dako Cytomation) for 30 min at RT.
Tissue sections were treated with 3%H2O22, blocked with 5% bovine serum albumin to prevent nonspecific binding, followed by incubation with primary antibody at 4°C overnight.
The sections were treated with 10% normal goat serum for 10 min to block non-specific binding.
After being treated with 3% hydrogen peroxide for 15 minutes to block the endogenous peroxidase, the sections were treated with normal goat serum confining liquid for 30 minutes to reduce non-specific binding and then rabbit polyclonal anti-IFIT2 (1 500, HPA003408, Sigma-Aldrich. Shanghai, China) was incubated the sections for 12 h at 4°C.
