Fig. 5: Elevated DMRT1 chromatin accessibility is associated with the extended differentiation potential of PSCs.

a Motif enrichment analysis of D-iPSC-specific enriched peaks in ATAC-seq (n = 270), performed using HOMER (Fisher’s exact test, two-sided). b Representative specific peaks in D-iPSCs and ATAC-qPCR analysis. Genomic regions containing DMRT1/6-binding motifs^{75, 76} and qPCR products are shown. Data are presented as means ± SD of biological duplicates. The mean ATAC-qPCR value of ESCs was defined as 1 (t-test, two-sided). c qRT-PCR analysis of Dmrt1 expression in ESCs, S-iPSCs, and D-iPSCs. Data are presented as means ± SD of technical triplicates. The mean expression level of ESCs was defined as 1 (one-way ANOVA, two-sided). d qRT-PCR analysis for expression of Dmrt1 in S-/D-OSKM kidney tumors. Data are presented as means ± SD of biological triplicates. Relative expression levels to ESCs are shown (one-way ANOVA and Dunnett’s multiple-comparison test, two-sided). e Representative histological images and immunostaining for OCT4, DAZL, and DMRT1 of D-OSKM kidney tumors. Scale bars: 200 μm. f Schematic illustration of the genomic construct of Dmrt1-inducible ESCs. g Representative images and immunostaining for CDX2 in Dmrt1-inducible EBs on differentiation Day 8. Scale bars: 200 μm (left), 500 μm (right). h qRT-PCR analysis of Cdx2 expression during EB differentiation. Data are presented as means ± SD of biological triplicates. The mean expression level in ESCs on differentiation Day 0 was defined as 1. i Representative histological images of tumors developed after inoculation of Dmrt1-induced cells into the testis of 6-week-old C57BL/6 N mice. Dotted lines indicate the TGC cluster. Right panel represents quantification of TGC cluster area in tumors (control, n = 6; Dmrt1-induced cells, n = 6). Data are presented as means ± SD of biologically independent samples. Scale bars: 500 μm (Mann–Whitney test, two-sided).