Figure 7.

Effects of Gln supplementation on epithelial regeneration and ISC differentiation in vivo and in vitro

(A) Quantification of apoptotic cells per villus top (n = 6 mice, means ± SEM; ^∗p < 0.05, ^∗∗p < 0.01; one-way ANOVA, results of 3 independent experiments).

(B) Quantification of cell migration after 24 h EdU labeling. EdU cell migration distance was used as an absolute measure of distance from the crypt bottom to the cell that had migrated the furthest (n = 4 mice, means ± SEM, ^∗∗p < 0.01, one-way ANOVA, results of 3 independent experiments).

(C) Representative confocal images of cell migration in suckling and weaned mice with 24 h EdU labeling. Scale bars, 100 μm.

(D) Representative images of immunofluorescence represent the overlap of positive signal (green) and nuclear signal (blue). Scale bars, 20 μm.

(E) Relative mRNA levels of transit-amplifying cells Prom1 and Zfp652, enterocyte marker Vill, Paneth cells markers Lysozyme and Mmp7, enteroendocrine Chga and Globet cells marker Muc2 in enteroids were analyzed by qPCR (n = 8, 8 wells of enteroids per group, means ± SEM; ^∗p < 0.05, ^∗∗p < 0.01; one-way ANOVA, results of 3 independent experiments).

(F–H) The number of goblet cells (F), enteroendocrine cells (G), and Paneth cells (H) per crypt-villus were quantified (n = 6 mice, means ± SEM, ^∗p < 0.05, one-way ANOVA, results of 3 independent experiments).

(I) Representative images of goblet cells. Scale bars, 200 μm. (J and K) Representative immunofluorescence images of lysozyme (red), CHGA (red), and DAPI (blue) in the jejunum. Scale bars, 100 μm.