FIGURE 3.

AF9 suppresses cancer initiation and progression in AOM/DSS induced CRC model. (A) Scheme of the Af9 gene locus and related alleles. The Af9‐floxed alleles have two loxP sites (red triangles) flanking the ninth exon (grey boxes). Mice with Af9‐floxed alleles were crossed with a Cre line to generate the deleted Af9 allele. (B) Western blot analysis of colon tissues from AF9^IEC+/+ and AF9^IEC‐/− mice using indicated antibodies. (C) Genotypes were determined by PCR using genomic DNA from intestinal epithelial cells. (D) AF9^{IEC‐ /−} was identified by IHC using antibody against AF9. (E) Body weight of AF9^IEC+/+ and AF9^IEC‐/− mice was measured after treatment with DSS. Body weight changes in the mice during three cycles of 3% DSS treatment. (F and G) The mice were euthanised to measure colon length (n = 6) and tumour burden (n = 6) on Day 36. (H) Hematoxylin and eosin (H&E)‐stained sections of middle–distal colon tissue. Arrows, infiltration of immune cell and epithelial cell damage. (I) Representative images of ZO‐1 staining in colon sections of mice treated with DSS. (J) Kaplan–Meier plot showing the survival of AF9^IEC+/+ and AF9^IEC‐/− mice after AMO/DSS treatment (*p < .05, **p < .01).