Fig. 6: Ehf/Cdx1 deletion increases susceptibility of mice to AOM/DSS-induced colorectal tumourigenesis.
A, B Confirmation of (A) Ehf and (B) Cdx1 knockout in the colonic epithelium of Ehf and Cdx1 knockout mice. Colonic epithelial cells were isolated from 6-week-old WT, EhfKO, Cdx1KO and EhfKO;Cdx1KO mice and Ehf and Cdx1 mRNA determined by q-RT-PCR. Values shown are mean ± SEM of n = 5 mice. C, D Body weights of 8-week-old (C) male and (D) female WT (n = 7 and n = 7 for male and female cohorts, respectively), EhfKO (n = 9 and n = 7), Cdx1KO (n = 7 and n = 10) and EhfKO;Cdx1KO (n = 6 and n = 7) mice. Values shown are mean ± SEM. E Average colitis disease score of WT (n = 8), EhfKO (n = 8), Cdx1KO (n = 9) and EhfKO;Cdx1KO (n = 9) mice following the second round of DSS treatment. Values shown are mean ± SEM. F Representative endoscopy images from WT, EhfKO, Cdx1KO and EhfKO;Cdx1KO mice showing inflammation (*) and colon tumour formation (arrows) in Cdx1KO and EhfKO;Cdx1KO mice. Endoscopies were performed in 13-week-old mice after receiving 2 injections of AOM and 2 rounds of DSS. G–I Quantitation of (G) tumour number, H overall tumour burden and (I) colon length in WT (n = 7), EhfKO (n = 4), Cdx1KO (n = 9) and EhfKO;Cdx1KO (n = 8)mice at endpoint. Values shown are mean ± SEM, with some EhfKO having to be euthanized for non-tumour related phenotypes (genital abscess). J Quantitation of KRT20 staining in tumours derived from WT (n = 5), EhfKO (n = 4), Cdx1KO (n = 5) and EhfKO;Cdx1KO (n = 4) mice. Values shown are mean ± SEM. K Representative immunohistochemistry images in the colon from WT, EhfKO, Cdx1KO and EhfKO;Cdx1KO mice stained with anti-KRT20 antibody. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, one-way ANOVA with Tukey’s post hoc test.
