Anti gapdh antibody

B cells were harvested and lysed on ice for 15 min in RIPA lysis buffer (Pulilai Gene Technology, Beijing) supplemented with a protease inhibitor cocktail (MedChemExpress, USA). Cell lysates (20 µg per sample) were denatured and subjected to SDS-PAGE (10%), followed by transfer to polyvinylidene fluoride (PVDF) membranes. The membranes were blocked with 5% skim milk and subsequently immunoblotted with anti-B4GALT1 antibody (1:250 dilution; Abcam), anti-GLB1 antibody (1:1000 dilution; Abcam), anti-CD75/ST6GAL1 antibody (1:1,000 dilution; Abcam), anti-sialidase-1 (NEU1) antibody (1:2,000 dilution; Abcam), and anti-GAPDH antibody (1:2,000 dilution, TransGen Biotech, China) overnight at 4°C. HRP-conjugated secondary antibodies were added for incubation, and then blots were developed with SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, USA). ImageJ software (developed at the National Institutes of Health) was used to quantify the relative staining intensity. Each experiment was conducted with at least three batches from different donors.

The following primary antibodies were used: PRRSV N protein antibody (JNT, China; no. JN0401), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH) antibody (TransGen Biotech, China; no. HC301), rabbit anti-HA (Cell Signaling Technology, USA; no. 3724 T), rabbit anti-Flag (Abmart, China; no. T20008M), mouse anti-HA (Abmart; no. M20003M), mouse anti-Flag (Beyotime, China; no. AF519), rabbit anti-GFP (Proteintech; no. 50430-2-AP), mouse anti-GFP (Proteintech; no. 66002-Ig), rabbit anti-β-actin (Proteintech; no. 81115-1-RR) and mouse anti-dsRNA (Scicons, Hungary; no. 10010500). The anti-ZNF283 antibody was generated by immunizing rabbits with bacterially expressed full-length porcine ZNF283.