Figure 2.

Increases in villus height, crypt density, ISC number, and proportion of ISC in S-phase in DIO vs control mice. A, Representative hematoxylin and eosin stained photographs depicting crypt-villus architecture of intestinal epithelium from control and DIO mice. Images were taken at 10x magnification. Scale bar, 100 μm. B, Villus height in control and DIO mice. C, Crypt density quantified by number of crypts per mm submucosal circumference in control and DIO mice. D, Number of cells per crypt in control and DIO mice. E, Number of cells in S-phase measured by EdU positive staining per crypt. F, Representative images of crypt sections stained with DAPI (blue), GFP (Sox9-EGFP; green), and Chromogranin-A (ChgA; red). Sox9-EGFP^Low ISC (white arrows) and Sox9-EGFP^High cells (open triangles) were defined by intensity of EGFP staining. G, Quantification of Sox9-EGFP^Low and Sox9-EGFP^High cells. H, Representative images of crypt sections stained with DAPI, GFP, and EdU (red). Dual positive Sox9-EGFP^Low and EdU cells are denoted by the filled triangles. EdU: 5-ethynyl-2′-deoxyuridine. Images were taken at 63x magnification. Scale bar, 20 μm. I, Percentage of Sox9-EGFP^Low or Sox9-EGFP^High cells positive for EdU. J, Relative abundance of Sox9-EGFP^Low cells assayed by flow cytometry and expressed as the percentage of total cells that fall in Sox9-EGFP^Low gates. Data expressed as mean ± SEM. *, P < .05 DIO vs control, unpaired t test, n ≥ 5.