Intestinal Protein Expression Analysis

Relative protein levels of CuSOD, MnSOD, GPX1, GPX4, ZO-1, Claudin-1, Occludin, IL-2, IL-8, and IL-10 in the jejunum were determined using western blotting. Colon samples were collected and the protein expression of CuSOD, MnSOD, GPX1, GPX4, IL-2, IL-8 and IL-10 was determined [22 (link)]. The resultant signals were obtained using Quantity One software (Bio-Rad, Hercules, CA, USA). Primary antibodies were used as follows: CuSOD (1:50000; ab51254, Rabbit, Abcam, UK), MnSOD (1:1000; ab68155, Rabbit, Abcam, UK), GPX1 (1:1000; bs-3882R, Rabbit, Bioss, Beijing, China), GPX4 (1:1000; 14432-1-AP, Rabbit, Proteintech, Rosemont, IL, USA), ZO-1 (1:3000; 21773-1-AP, Rabbit, Proteintech, USA), Occludin (1:1000; ab167161, Rabbit, Abcam, UK), Claudin1 (1:500; ab15098, Rabbit, Abcam, UK), IL-2 (1:2000; ab92381, Rabbit, Abcam, UK), IL-8 (1:2000; ab110727, Rabbit, Abcam, UK), IL-10 (1:1000; 20850-1-AP, Rabbit, Proteintech, USA) and Actin (1:5000; 66009-1-Ig, Mouse, Proteintech, USA).