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. 2023 Jun 15;18(7):1451–1467. doi: 10.1016/j.stemcr.2023.05.012

Figure 5.

Figure 5

Gln supplementation augments the activity of ISCs in vitro

(A). Representative bright-field images of the stable-passaged enteroids grown for 72 h in medium containing 0, 2, or 5 mM Gln. Scale bars, 100 μm.

(B and C) Crypts domains and surface area of stable-passaged enteroids were measured (n = 12, 12 enteroids per condition were analyzed, means ± SD; ∗p < 0.05, ∗∗p < 0.01; one-way ANOVA, results of 4 independent experiments).

(D) Heat maps were generated based on levels of the stemness marker genes of ISC cells in vitro. The data represent fold-change differences relative to the 0 mM Gln group. Genes with a corresponding adjusted p value less than 0.05 were considered statistically significant (n = 4, 4 independent wells of enteroids, ∗p < 0.05, one-way ANOVA, results of 3 independent experiments).

(E) Representative images of enteroid immunofluorescence stained with OLFM4 (green) and DAPI (blue) at 72 h. Scale bars, 100 μm.

(F) Western blot analysis of intestinal stem cell marker OLFM4 protein expression (n = 4, 4 independent wells of enteroids per group, results of 3 independent experiments).