The protein levels of Claudin-2, OAS2 and OAS3 in colon tissues in each group were detected by Western blotting. In brie y, total proteins from colon tissues were obtained with RIPA lysis buffer (Beyotime, Shanghai, China) and protease inhibitor (Servicebio, China). After the determination of protein concentration, the protein was denatured by boiling water for 15 minutes. The primary antibodies claudin-2 (AF0128, A nity Biosciences, USA), OAS2 (DF12680, A nity Biosciences, USA), OAS3 (ER1914-27, HuaBio, China), and anti-β-actin (AF7018, A nity Biosciences, USA) were thinned and incubated overnight at 4 °C. We added HRP-labeled goat anti-rabbit secondary antibodies in a 1:3000 dilution, and incubated them at room temperature for 30 minutes.
Wash the membranes three times with PBST. The immunoreactive bands were detected with ECL Luminescent Solution (Servicebio, China).
TdT-mediated dUTP Nick-End Labeling (TUNEL) Assay Apoptosis of IECs was detected by TUNEL kit (Servicebio, China). In short, colon tissue sections were cultured in TUNEL reaction mixture. Then, it was stained with 4', 6-diamino-2-phenylindole (DAPI). After that, all samples were observed under the uorescence inverted microscope. Each sample was observed in three random elds.
Wash the membranes three times with PBST. The immunoreactive bands were detected with ECL Luminescent Solution (Servicebio, China).
TdT-mediated dUTP Nick-End Labeling (TUNEL) Assay Apoptosis of IECs was detected by TUNEL kit (Servicebio, China). In short, colon tissue sections were cultured in TUNEL reaction mixture. Then, it was stained with 4', 6-diamino-2-phenylindole (DAPI). After that, all samples were observed under the uorescence inverted microscope. Each sample was observed in three random elds.