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. 2020 Jun 15;136(12):1442–1455. doi: 10.1182/blood.2020005957

Figure 1.

Figure 1.

Loss of enteroendocrine L cells and its peptide GLP-2 upon GVHD induction. (A) Representative immunohistochemistry staining for GLP-2 (red) in colon sections. Shown are sections from untreated animals, mice undergoing syn-HCT on day 10, mice undergoing allo-HCT on day 10 and allo-HCT mice on day 10 conditioned with chemotherapy. Scale bars, 20 μm. (B) GLP-2+ cells quantification in untreated C57BL/6 (n = 5), untreated BALB/c mice (n = 10), TBI/syn-HCT BALB/c mice on day 10 (n = 10), chemotherapy/allo-HCT BALB/c mice on day 10 (n = 10), TBI/allo-HCT C57BL/6 mice on day 10 (n = 10) and TBI/allo-HCT BALB/c mice on day 10 (n = 10). Results from 1 or 2 independent experiment(s) are shown. (C) Relative gene expression of the L-cell marker Cldn4 mRNA in the small intestine of untreated BALB/c (n = 10), syn-HCT on day 10 (n = 10), and allo-HCT BALB/c mice on day 5 (n = 10). Gene expression was normalized to β-actin used as reference gene. Results are derived from 2 independent experiments. (D) Relative gene expression of the GLP-2R (receptor) in the large intestine in BALB/c mice under GVHD conditions euthanized at day 2 (n = 6), day 4 (n = 6), day 7 (n = 11), day 14 (n = 13) compared with untreated BALB/c mice (n = 10). Gene expression was normalized to β-actin used as reference gene. Results are derived from 2 independent experiments. (E) Representative western blot showing the GLP-2 protein levels in the small intestine of BALB/c mice undergoing TBI-based allo-HCT on day 10 in comparison with untreated mice. β-actin was used as loading control. (F) Quantification of GLP-2 total protein. Fold change was normalized to the β-actin levels and relative to the untreated controls (set as “1”). Results are from 2 independent experiments at 2 different time points, day 5 (n = 11) and day 10 (n = 8) after allogeneic HCT. (G) Representative western blots from the gastrointestinal tract (colon) of chemotherapy-based allo-HCT BALB/c mice on day 3 in comparison with untreated mice. (H) Quantification of GLP-2 protein levels in chemotherapy-based allo-HCT BALB/c mice (n = 10) vs the untreated group (n = 8). Results are shown from 2 independent experiments. The P values in this figure were calculated using the unpaired, 2-sided Student t test.