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. 2025 Aug 28;5(1):100774. doi: 10.1016/j.gastha.2025.100774

Figure 1.

Figure 1

Generation of TM6SF2 E167K-edited iPSC-derived hepatocyte-like cells. (A) Schematic illustration of the gene-targeting strategy for the TM6SF2-rs58542926 variant using the MhAX method. Homozygous (KK), heterozygous (EK), and wild-type (EE) iPSC lines were generated from a wild-type human iPSC line. (B) Sanger sequencing was performed to examine whether the human iPSC clones were correctly targeted. (C) Phase contrast images of genome-edited HLCs are shown. (D) Genome-edited HLCs were subjected to immunostaining with anti-ALB, anti-ZO1 and anti-CK18 antibodies. Nuclei were counterstained with Hoechst 33,342. (E and F) Gene expression levels of hepatic marker genes (ALB, ASGR1, HNF4A, CEBPA, and NR1H3) and VLDL assembly-related genes (APOB, MTTP, TM6SF2, and LPCAT3) in genome edited HLCs were examined by qRT-PCR (∗P < .05; Tukey’s post hoc tests). (G) Protein levels of HNF4α, FOXA2, Albumin, and α-tubulin in genome-edited HLCs were examined by western blotting. (H) Amounts of ALB secretion in genome-edited HLCs and PHHs were examined by ELISA (Tukey’s post hoc tests). All data represent mean ± SD. n.s., not significant.