Anti gnaz

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).

After the pre-cleaning process, whole-cell lysates (0.5 mg per reaction) were incubated overnight (4 °C) with 1–2 μg of immunocapturing antibody, which included anti-ADAM17 (Bioss Antibodies, Woburn, MA, catalog no. bs-4236R) and anti-GNAZ (Genetex, Irvine, CA, catalog no. GTX11444). The next day, protein G magnetic beads (Merck Millipore, Burlington, MA) were added and incubated for another 2 h. All the incubations were performed on a rotating shaker at a 4 °C refrigerator. Finally, the immunocomplexes were washed three times and separated using SDS-PAGE. Immunoblotting was performed according to the standard procedures.