Fig. 1. Characterization of ALDH2*1/*1-hiPSCs and ALDH2*1/*2-hiPSCs. (A) Schematic representation of hiPSC generation. (B) DNA sequencing results of ALDH2*1/*1- and ALDH2*1/*2-hiPSCs confirming ALDH2 mutation. (C) Measurement of ALDH2 activity in ALDH2*1/*1-hiPSCs (n=3) and ALDH2*1/*2-hiPSCs (n=3). (D) Bright field microscope images and alkaline phosphatase (AP) staining of ALDH2*1/*1- and ALDH2*1/*2-hiPSCs. Scale bar=200 μm. (E, F) Gene expression of pluripotency markers in ALDH2*1/*1-hiPSCs (n=3) and ALDH2*1/*2-hiPSCs (n=3), determined by RT-PCR and qRT-PCR. GAPDH was used as a loading control. Relative gene expression values were calculated using the delta-delta Ct (2^(-ΔΔCt)) method. (G) Immunocytochemical staining for pluripotency markers OCT4, SOX2, KLF4, TRA-1-60, TRA-1-81, and SSEA4. Scale bar=100 μm. Data are presented as the mean±SEM. ^##p<0.01 (t-test) indicates statistical significance. PBMC: peripheral blood mononuclear cell, ALDH2: acetaldehyde dehydrogenase 2, hiPSCs: human induced pluripotent stem cells, RT-PCR: real-time polymerase chain reaction, qRT-PCR: quantitative RT-PCR.