Fig. 6: Induction of PrE by SALL4^delN12 alone.

a Immunofluorescence staining shows the iPrE shape clone is GATA4 and LAMA1 dual positive induced by SALL4^WT and SALL4^delN12 at day 11, scale bar = 250 μm. b Pictures show the cell morphology change along iPrE induction by SALL4^WT and SALL4^delN12. c Bar plot for GATA4 positive iPrE colonies numbers under BMP4− conditions at Day 11, data are mean ± s.d., two-sided, unpaired t-test; n = 3 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001. d Whole well screening photograph of (c), scale bar = 5 mm. e and f Box plot shows PrE-related gene expression between SALL4^WT and SALL4^delN12 at different time points. A two-tailed Student’s t-test was performed for comparisons between classes (***p-value < 0.001; **p-value < 0.01; *p-value < 0.05; -p-value > 0.05) The middle lines of the boxes indicate the median, the outer edges represent the first and the third quartiles, and the whiskers indicate the 1.5 × interquartile range below the lower quartile and above the upper quartile. g Heatmap shows the differentially enriched motifs in the CO groups between SALL4^delN12 and SALL4^WT, as referenced in Supplementary Fig. 6j. h Line chart shows the average SALL4 binding signal profile between SALL4^delN12 and SALL4^WT at ±2k of TSS of PrE marker genes. i Box plot shows SALL4 binding signal differences between SALL4^delN12 and SALL4^WT at ±2k of TSS of PrE marker genes in (j). A two-tailed Student’s t-test was performed for comparisons between classes. “–“ means no significant difference (p-value = 0.05), The middle lines of the boxes indicate the median, the outer edges represent the first and the third quartiles, and the whiskers indicate the 1.5 × interquartile range below the lower quartile and above the upper quartile. j Line chart shows the average H3K27ac signal profile between SALL4^delN12 and SALL4^WT at ±2k of TSS of PrE marker genes. k Box plot shows the H3K27ac signal differences between SALL4^delN12 and SALL4^WT at ±2k of TSS of PrE marker genes in (i). A two-tailed Student’s t-test was performed for comparisons between classes. (***p-value < 0.001). The middle lines of the boxes indicate the median, the outer edges represent the first and the third quartiles, and the whiskers indicate the 1.5 × interquartile range below the lower quartile and above the upper quartile.