Measurement of Colonic Phenotype

At the end of the animal experiment, mice were sacrificed by cervical dislocation under anesthesia using isoflurane. After being isolated from the abdomen, the colon tissue was cut longitudinally and flushed with the 4% cold PBS wash buffer to clean up feces and bacteria. We dry the colon with sterile gauze and measure the length and weight of the colon. Then the colonic tumors are counted, weighed, and volume measured. The proximal colon was stored in germ-free 1.5 ml centrifuge tubes, frozen in −80 C refrigerator; the distal colon was fixed in 4% paraformaldehyde overnight and embedded in paraffin routine H&E histopathologic examination. The whole tissue collection process is carried out at low temperature (0–4°C), and colon tissues' characterization was photographed.