Fig. 3. Characterization of InS^exp derived from immature and mature 3D HIOs. (A, B) Effect of CHIR99021 on InS^exp derived from immature and mature HIOs. Bright field images of InS^exp (A) and cell number increase of the InS^exp after day 14 (B). White scale bars, 500 μm. Yellow scale bars, 250 μm. Data represent means±SEM (n=4). (C) Relative gene expression of intestinal epithelial cell markers and WNT target genes in InS^exp derived from immature and mature HIOs. (D, E) Immunofluorescence analysis of the intestinal epithelial cell markers and WNT target genes in InS^exp derived from immature and mature HIOs. White scale bars, 50 μm. *p<0.05, **p<0.01, ***p<0.001 using two-tailed t-test.