Figure 2.

Determination of the regulatory elements underlying cell-fate bifurcation of germline and non-germline lineages

(A) Venn diagram showing the top 10k common and specific peaks in hESCs, 4i cells, and day 1 cells.

(B) Heatmap showing TF motifs significantly enriched in the common and specific peaks in hESCs, 4i cells, and day 1 cells defined in (A).

(C) The top binding motifs enriched in day 1-specific peaks.

(D) Venn diagram showing the top 10k common and specific peaks in day 1, D2DP, and D2N cells.

(E) Heatmap showing TF motifs significantly enriched for common and specific peaks in day 1, D2DP, and D2N cells defined in (D).

(F) The top binding motifs enriched in D2DP-specific and D2N-specific peaks.

(G) Failed-to-open (FO) and failed-to-close (FC) peaks for D2N, D4N, D4SP, and D6N cells compared with D6DP cells are shown. These peaks are derived from peaks open in 4i cells yet closed in D6DP cells or peaks closed in 4i cells yet open in D6DP cells. (H) Heatmap showing TF motifs significantly enriched in FO and FC peaks in D2N, D4N, D4SP, and D6N cells.

(I) Gene expression of TEAD, GATA family, and AP1 family TFs among DP and N cells as well as 7-week hPGCs (Irie et al., 2015).

(J) Immunofluorescence of JUN, GATA4, and SOX17 in EBs from day 1 to day 6, Scale bar, 50 μm. The dotted boxes enclose representative zoomed images.

(K and L) Quantification of relative fluorescence intensity of GATA4 (K) or JUN (L) for SOX17-positive and -negative cells measured by the ImageJ software. Eight slides of immunostaining from three independent experiments were used. Two-tailed Student's t test was performed, ^∗∗∗∗p < 0.0001.