Fig. 1: Genotoxic stress-induced stem cell maldifferentiation (GSMD) occurs in hESCs via TGF-β signaling.

a Schematic of the workflow. b Stemness in hESCs was evaluated using FITC-conjugated rBC2LCN in the presence or absence of 5-FU for 24 h. Images are representative of n = 3 independent experiments with similar results. c Heatmap displaying the expression of representative mRNAs involved in pluripotency (n = 3 biologically independent samples). d Radar charts depicting the expression of typical triploblastic genes in hESCs after 5-FU treatment. Data are presented as mean ± SEM from n = 3 biologically independent samples. (No statistical test was performed.). e Genes differentially expressed in a p53-dependent manner from RNA-seq (n = 3 biologically independent experiments). f p53-dependent upregulated genes (327) overlapping with cell development-related genes (GO:0048468). The 327 and 2140 genes identified in VENNY 2.1 were subjected to GO analysis (panels g, h). g GO analysis of the 327 upregulated developmental genes using BioPlanet 2019. Statistical significance was determined by two-sided Fisher’s exact test with Benjamini–Hochberg (BH) correction; exact p-values are shown. h GO analysis of the 327 upregulated developmental genes using ARCHS4 Tissues. Statistical significance was determined by two-sided Fisher’s exact test with BH correction; exact p-values are shown. i TGF-β signaling was evaluated by immunostaining for SMAD3 and FITC–rBC2LCN under 5-FU, ADR, or DNR. Images are representative of n = 4 independent experiments with similar results. Scale bars, 50 μm.