Fig. 5. Selected m⁶A-disruptive mutations regulate human endodermal specification through m⁶A.

a mRNA levels of DE marker genes SOX17, FOXA2, CXCR4, GATA4, GATA6, and HNF1B in cultures derived from WT, SOX2-mut, SDHAF1-mut, and ADM-mut as well as WT hESCs at day 3 of DE differentiation (n = 6 biologically independent samples). P-values were calculated vs. WT (two-tailed Student’s t-test). b, c Representative (b) and quantitative (c) flow cytometry analyses of CXCR4 expression in cultures derived from WT, SOX2-mut, SDHAF1-mut, and ADM-mut hESCs at day 3 of DE differentiation (n = 6 biologically independent samples). P-values were calculated vs. WT (one-way ANOVA with Tukey’s post hoc test). d, e Measurement of m⁶A levels of SDHAF1-c.*76 (d) and ADM-c.*68 (e) after TRME editor induced site-specific m⁶A demethylation by SELECT assay (n = 6 biologically independent samples). NT non-targeting, Dox doxycycline (one-way ANOVA with Tukey’s post hoc test). f, g mRNA expression levels of SDHAF1 (f) and ADM (g) on day 2 of DE differentiation after TRME editor induced site-specific m⁶A demethylation (n = 6 biologically independent samples). NT non-targeting, Dox doxycycline (one-way ANOVA with Tukey’s post hoc test). h, i Representative (h) and quantitative (i) flow cytometry analyses of CXCR4 expression in cultures derived from hESCs with or without induced site-specific m⁶A demethylation by TRME editor at day 3 of DE differentiation (n = 6 biologically independent samples). NT non-targeting, Dox doxycycline (one-way ANOVA with Tukey’s post hoc test). Data of all relevant panels are presented as means ± SD. Source data are provided as a Source Data file.