Figure 3. Vancomycin treatment of WT mice depleted DCA in the wounds and impaired crypt regeneration in a PGE2-dependent manner.

(A) Wound tissue from WT mice was collected on days 4 and 6 post-injury and analyzed for DCA levels by mass spectrometry (n=17–27 wounds/ group from 6–8 mice per group). Significance was determined by one-way ANOVA and TUKEY’s post hoc test: *p<0.05. Sham=colonic tissue from mice that underwent endoscopy but not biopsy injury. (B) Schematic representation of the procedures and analysis (in red text) measured on days 6 and 12 in C-F. See also Figure S4. (C) Wounds from mice treated with either vehicle or vancomycin were analyzed for DCA at day 6 post-injury, ND= not detected. (D) Wound PGE2 levels at day 6 post-injury from vehicle and vancomycin treated mice (n=19–21 wounds/group from 7–8 mice/group). Significance was determined by unpaired Student’s t test: **p<0.01. (E) Representative images of H&E stained sections (Bars=100µm) and (F) wound bed length at day 12 post-injury from indicated groups of mice (n=16–19 wounds/group from 6–9 mice/group). Significance was determined by unpaired Student’s t test: ***p<0.001. (G-H) Mice treated with vancomycin were biopsy injured, treated with either vehicle or NS-398 (twice daily from days 4–10 post-injury) and wounds were analyzed. (G) Representative images of H&E stained sections (Bars=100 µm) and (H) wound bed length at day 12 post-injury (n=14–19 wounds from 6–8 mice/group). Significance was determined by unpaired Student’s t test: ****p<0.0001. All values in A, C, D, F and H are displayed as mean ± SEM. See also Figure S4.