Figure 4. Esrp1^Triaka modulates the course of experimental intestinal immunopathology.

(A) WT (n = 7) and Triaka (n = 4) mice were challenged with 2% DSS in the drinking water for 7 days. Weight loss was measured daily (left panel). One representative out of four different experiments is shown. Representative H&E slides from colonic sections illustrate the degree of pathology in the different groups 7 days after the start of DSS treatment (right panel). Scale bars: 200 µm. (B) Colon length was measured in DSS-treated mice (n = 7–12 mice per group). (C) A miniature forceps was used to induce injuries in the colonic mucosa of the indicated groups of mice. Wound-healing was monitored by colonoscopy. Representative pictures (left panel) and quantification of wound-healing over time (right panel) are shown. Right panel represents pooled data from four independent experiments (n = total of 17 wounds from 8 to 9 mice, per group). (D) WT and Triaka mice were treated with AOM/DSS and sacrificed. Number of tumors, (E) tumor size and (F) the highest tumor grade per mouse are shown, assessed 70 days after the initial AOM injection. For (D), (E) and (F), one representative experiment of two is shown (n = 8–9 mice per group). (G) WT and Triaka mice were treated with 2% DSS in the drinking water for 3 days and Ki-67 staining was performed. Representative pictures are shown and (H) Ki-67-positive cells were quantified (n = 9–10 mice per group). Statistics: (A) and (C) Two-way ANOVA with Bonferroni post-test, (B) and (H) Student's t test, (D) Mann-Whitney test and (E) Student’s t test with Welch’s correction. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Figure 4—figure supplement 1. Triaka mice show increased susceptibility to experimental colitis.

(A) WT and Triaka mice were treated with 2% DSS water and disease activity score was assessed at the indicated time points (n = 4–7 mice per group). (B) Colitis score was assessed by histological analysis at the indicated time points after start of DSS treatment (n = 4–5 mice per group). (C) Water consumption of single-housed mice of the indicated genotypes was measured daily during 7 days and averaged (n = 6–7 mice per group). (D) Mice were given 3 cycles of DSS alternated with regular water, as indicated, and weight loss was measured (n = 5 mice per group). Data represent: Means ± standard error of the mean from (A) one representative experiment repeated three times. Statistics: (A) and (D) Two-way ANOVA with Bonferroni post-test. (B) Student's t test. *p<0.05; **p<0.01; ***p<0.001.

Figure 4—figure supplement 2. More aggressive molecular signature in Triaka intestinal tumors.

WT and Triaka mice were treated with AOM/DSS. Seventy days after the first AOM injection, tumors or adjacent tumor-free (healthy) colonic tissue were homogenized and the indicated proteins were measured by Multiplexing LASER Bead Technology. n = 5–8 mice per group. Statistics: Student's t test. *p<0.05; **p<0.01.

Figure 4—figure supplement 3. Partial EMT signature in Triaka cIECs.

Quantitative PCR was applied to measure transcript levels of (A) Cdh1, (B) Zeb1, (C) Zeb2, and (D) Esrp1 in WT and Triaka cIECs, after normalization to Gapdh expression. n = 9–14 mice per group. Statistics: Student’s t test. *p<0.05, **p<0.01.