Rabbit anti claudin 1

Western blot analysis was performed to determine the jejunal mucosa protein expressions of occludin, claudin-1, ZO-1, p–NF–κB p65, NF-κB p65, p-IκBα and COX-2, as previously described (Xun et al., 2021 ). The primary antibodies were as follows: rabbit anti-β-actin (Boster, Wuhan, China) in the dilution of 1:2,000, rabbit anti-occludin (Proteintech, Wuhan, China) diluted at 1:1,000, rabbit anti-claudin1 (Bioss, Beijing, China) in 1:1,000 dilution, rabbit anti-ZO-1 (Proteintech, Wuhan, China) in 1:1,000 dilution, rabbit anti-COX-2 (Proteintech, Wuhan, China) in 1:1,000 dilution, rabbit anti–NF–κB p65 (Cell Signalling Technology, Danvers, MA, USA) at 1:1,000, rabbit anti-phospho–NF–κB p65 (Cell Signalling Technology, Danvers, MA, USA) in 1:1,000 dilution, and rabbit anti-phospho-IκBα (Cell Signalling Technology, Danvers, MA, USA) at 1:1,000 dilution. The secondary antibody was horseradish peroxidase (HRP)-conjugated anti-rabbit antibody (Cell Signalling Technology, Danvers, MA, USA) and it was diluted at 1:2,000. Protein abundance was normalized with housekeeping protein β-actin and stated as fold change. The results were communicated relative to the control piglet's levels.