Figure 2.

Intestinal Differentiation Capacity of PEPT1-KO Human iPSCs

(A) The procedure for intestinal differentiation from human iPSCs is shown. (B) The gene expression levels of intestinal markers (VIL1, ISX, and SI) and PEPT1 in WT-iPSCs and PEPT1-KO iPSC-IECs were examined by real-time RT-PCR analysis. The gene expression levels in the WT iPSC-IECs were taken as 1.0. Data represent the means ± SD. Statistical analyses were performed using the unpaired two-tailed Student’s t test (***p < 0.001). (C) The protein expression levels of PEPT1 and β-actin in WT iPSC-IECs and PEPT1-KO iPSC-IECs were examined by western blotting analysis. (D) The percentages of VIL1-positive cells and SI-positive cells in WT iPSC-IECs and PEPT1-KO iPSC-IECs were measured by FACS analysis. Data are presented as means ± SD (n ≥ 6, technical replicate). (E) Immunostaining analysis of VIL1 (red) was performed in WT iPSC-IECs and SLC15A1-KO iPSC-IECs. Nuclei were stained with DAPI (blue). Scale bars represent 50 μm. (F) The protein expression levels of VIL1 and β-actin in WT iPSC-IECs and PEPT1-KO iPSC-IECs were examined by western blotting analysis.