Hematoxylin and eosin (H&E) staining

Colon histopathology was evaluated by H&E staining. Briefly, ~24 h after fixing with neutral formalin, the mice colonic tissues were dehydrated using a graded series of alcohol solutions and embedded in paraffin. The embedded colonic tissues were sliced into sections of a thickness of 5 µm. The sections were placed in an oven and heated at 65˚C for 30 min. Next, the sections were stained with hematoxylin (cat. no. C0107; Beyotime Institute of Biotechnology) and eosin (cat. no. C0109; Beyotime Institute of Biotechnology), as reported elsewhere (26). Finally, the sections were sealed with neutral resin and scanned using a light microscope (Ax70; Olympus Corporation) (magnification, x400) with CellSens Standard v.1.6 software (Olympus Corporation).