Figure 3.

Lgr5+ cells persist in oxazolone colitis. Oxazolone (OXA) was intrarectally distilled into BALB/c mice. Outcomes were compared against ethanol- or PBS-treated mice (n ≥ 5 male mice per group). Body weights (A and E), colonic size (B and F), pathology score (C and G), and hematoxylin-eosin (H&E)-stained images (D and H) of mice treated with oxazolone, ethanol (vehicle), or PBS for 1 day (A–D) or for 5 days (E–H). Oxazolone-treated mice exhibit significant exacerbation of injury vs. ethanol-treated animals. Histological images showing the localization of Lgr5 (I), Ephb3 (J), Lrig1 (K), and Ly6a (L), through in situ hybridization (ISH), at day 1 or day 5 of treatment with either saline, ethanol vehicle, or oxazolone. A similar crypt-to-crypt variability in retention of Lgr5 staining was observed, as in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-treated animals. The expression of Ly6a was highest in oxazolone-treated day 1 samples). M: densitometry of the staining signal of Lgr5, Ephb3, Lrig1, and Ly6a in 10 contiguous representative crypts labeled by ISH. The overall pattern was consistent with retention of Lgr5 expression during oxazolone-induced colitis. Scale bars: D and H: 100 µm; I–L: 25 µm. Statistics: *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Pairwise two-tailed t tests with Benjamini-Hochberg multiple testing correction. Error bars: means ± SE.