Fig. 2

Generation of the hIECs model for drug absorption. a Schematic of the hIECs model establishment from iPSCs. b Bright-field images of normal (left) and patient-derived hIECs (right). Scale bar, 200 μm. c mRNA expression of intestinal cell type markers: CD44, SOX9, LGR5 (stem cell related), VIL1 (enterocyte), MUC2, MUC13 (goblet), LYZ (Paneth), CHGA (enteroendocrine), and ECAD (epithelial). d Immunofluorescence staining of intestinal cell-type markers and E-cadherin (top) with an enlarged view of the left panel (white box) and fluorescence intensity quantification (bottom). Scale bar, 200 μm. e mRNA expression of phase I/II drug metabolism markers. f CPR activity in normal and patient-derived hIECs. g TEER values of hIECs models and Caco-2 cells. h Immunofluorescence of CLDN1 and ZO-1. Scale bar, 200 μm. i Basolateral dextran-FITC uptake (4 kDa and 40 kDa) for passive permeability analysis. j Schematic (left) and mRNA expression (right) of intestinal influx/efflux transporters. A schematic illustration was created using Mind the Graph. k H&E staining (left) and immunofluorescence (right) of apical (VIL1, P-gp, PEPT1) and basolateral (Na⁺/K⁺ ATPase) markers. Nuclei stained with DAPI or Hoechst. Scale bar, 200 μm. Data are shown as mean ± SEM (n ≥ 4 per group). Statistical analysis for multiple groups (qPCR, IF, CPR activity, and TEER) was performed using one-way ANOVA with Tukey’s test for post hoc analysis, and data for IF was performed using a two-tailed unpaired Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ns: not significant