Extended Data Fig. 7: Differentiation of ESCs in vitro to generate distinct in vivo-like populations.

(a) Violin plots represent gene expression of the four most common genes used to purify DE in vitro. Two-sided Mann-Whitney U was used to determine the significance between VE clusters (AVE, EmVE, ExVE1 and ExVE2) and DE clusters (DE1 and DE2); *=<0.05; **=<0.01; ***=<0.001; ns=non-significant. (b) FACS plots showing the sorting strategy for isolating specific cell populations at various stages of 2D-ESC differentiation using the Gsc-GFP/Hhex-RedStar double reporter. (c) Sample representation within each differentiation protocol. Samples distribute evenly across the appropriate clusters. (d) Expression of known lineage markers agrees with the assigned cluster identities. DE- definitive endoderm, PrE – primitive endoderm, PS- primitive streak, VE –visceral endoderm.