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Fig. 4. Involvement of Activin/Nodal, PI3-kinase and MAPK signaling pathways
in β-catenin-induced mesoderm and the anterior PS
differentiation. (A,C) Immunoblot analysis of total cell lysates
from 
Nβ-cateninER cells. Cells were cultured for 3 days with or
without 4OHT, Noggin (Nog; 250 ng/ml), Activin A (25 ng/ml) or BMP4 (10
ng/ml). Cell lysates were subjected to SDS-PAGE, and probed with specific
antibodies as indicated. (B) The Nβ-cateninER cells were
cultured with 4OHT and Noggin in the presence or absence of SB431542 (SB) for
3 days, and immunostained with anti-N-cadherin and FOXA2 antibodies. Scale
bar: 100 µm. (D) Localization of β-catenin in
Nβ-cateninER cells. Cells were cultured with or without 4OHT and
Noggin, and subjected to immunostaining with anti-β-catenin (total) and
anti-ER antibodies (to recognize exogenous Nβ-cateninER). Nuclei
were counterstained with DAPI. Scale bar: 50 µm.
