Histopathological and Immunohistochemistry analysis
Formalin-fixed mouse colon tissues were embedded in paraffin wax and cut into 3μM thick sections for histopathological and immunohistochemical examination. For histopathology, sections were dewaxed and hydrated, stained with hematoxylin (Sigma, USA) and eosin (Solarbio), and photographed with a Leica microscope to evaluate the morphology. Immunohistochemical analyses were performed for iNOS, F4/80, Notch1 and hes1. Colon sections were dewaxed and hydrated, repaired with microwave in PH 6, 0.1M sodium citrate buffer, washed with PBS for three times, immunohistochemical pen drew circles on the sections, and endogenous catalase inhibitors were dropped in the circles for 20min, and the sections were sealed with the serum working liquid in the ABC kit for 20min. Sections were incubated overnight with anti-iNOS, anti-F4/80, anti-Notch1 and anti-Hes1 antibodies at 4℃. Biotin-labeled secondary antibody was incubated at 37℃ for 30 minutes, peroxidase (DAB) was stained for 5 minutes, the positive area was stained brown, and finally stained with hematoxylin, dehydrated, transparent and sealed. Images were captured using a Leica microscope.