FIG. 2.

In three-dimensional culture, Geminin promotes epithelial-to-mesenchymal transition (EMT) and mesendodermal differentiation. (A) Immunohistochemical localization of E-Cadherin in a control (C) cell line and in Geminin (Gem) ESC grown without Dox as embryoid bodies (EBs) for 4 days, followed by 2 days in adherent culture. Compared to controls that express high levels of E-Cadherin (a), Geminin overexpression dramatically downregulated the expression of E-Cadherin protein (b), a hallmark of cells that have undergone EMT. Control EB have attached, but cells have not yet migrated from them as illustrated by Hoechst staining (c), unlike Geminin-overexpressing EBs that attach, and cells migrate rapidly from the EB (d). (e) and (f) are overlays of ac and bd. Scale bar=200 μM. (B) Localization of cell-type-restricted proteins in control (C) and Geminin (Gem) ESC grown as in (A) above. Geminin overexpression decreased the expression of the pluripotency marker (Oct3/4; a, b) and of a neural precursor marker (Sox3; c, d), but increased expression of a mesodermal marker (Brachyury; e, f) and a marker of the endoderm lineage (Foxa2; g, h). The nuclei were stained with Hoechst 33258 (blue). Scale bar=200 μM. Color images available online at www.liebertpub.com/scd