Anti tjp1

Confluent cells were conditioned in a blue light exposure chamber at indicative illuminances or time intervals. Next, the protein expression level was detected by western blotting, using a previously described protocol [22 (link)]. The antibodies used in this study were as follows: anti-CLDN5 (Abcam, Cambridge, UK), anti-ADAM17, anti-ACTB (Sigma-Aldrich, Chicago, IL), anti-ADAM17 (phospho T735) (Cusabio, Houston, TX), anti-GNAZ (Genetex, Irvine, CA), anti-OCLN (Proteintech, Chicago, IL), and anti-TJP1 (Invitrogen, Waltham, MA, USA).

The standard protocol of western blot was performed according to our previous reports.^{26 (link),28 (link)} Briefly, the concentration of total protein from segments of small intestine (ileum) was measured with a BCA protein assay kit. 20 µg of protein was fractionated in 12% SDS-polyacrylamide gels and transferred onto polyvinylidene difluoride membranes. The membranes were blocked with 5% freshly prepared milk-TBST for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: anti-Ocln (1:2000; Abcam, Cambridge, USA), anti-Tjp1 (1:1000; Invitrogen, Carlsbad, CA) and anti-Cldn4 (1:3000; Abcam, Cambridge, USA). The membranes were then washed three times for 10 min in TBST and incubated with secondary antibody conjugated with HRP for 2 h. Finally, the membranes were subjected to a chemiluminescence detection system and exposed to a photographic film. Immunoreactsive bands were quantified using Image J software and bands were normalized with β-actin.