Figure 6.

POUV Proteins Support Epithelialization and Antagonize EMT

(A) The diagram shows rescue of ZHBTc4 ESCs with different FLAG-tagged POUV proteins. POUV proteins were introduced into these cells in the presence of tetracycline (Tet), and cell lines expressing similar and Oct4-like levels of POUV proteins were selected for further analysis. See also Figure S5A.

(B–C) Downregulation of E-cadherin in response to Oct4 depletion in ZHBTc4 ESCs. ZHBTc4 ESCs were plated in self-renewing conditions in either the presence or absence of Tet and assessed by flow cytometry (B) or immunofluorescence (C). The scale bar represents 50 μm.

(D–E) POUV- and, particularly, Xlpou25-rescued cell lines express localized E-cadherin. FLAG-POUV cell lines were cultured under self-renewing conditions and assessed by flow cytometry (D) or immunofluorescence (E). The scale bar represents 50 μm.

(F) POUV- and, particularly, Xlpou25-rescued cell lines maintain normal epithelial morphology during mesoderm and endoderm differentiation. Immunofluorescence on cultures differentiated for 4 days. The scale bar represents 50 μm.

(G) Xlpou25-supported ESCs are particularly resistant to the induction of EMT. ESC lines rescued by the indicated POUV protein were plated on fibronectin to stimulate EMT. Morphology and expression of E-cadherin and p120-catenin were assessed by immunofluorescence microscopy.

See also Figure S5.