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Involvement of Activin/Nodal, PI3-kinase and MAPK signaling pathways in β-catenin-induced mesoderm and the anterior PS differentiation. 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 ΔNβ-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 ΔNβ-cateninER cells were cultured with 4OHT and Noggin in the presence or absence of SB (SB) for 3 days, and immunostained with anti-N-cadherin and FOXA2 antibodies. Scale bar: 100 μm. (D) Localization of β-catenin inΔ 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 ΔNβ-cateninER). Nuclei were counterstained with DAPI. Scale bar: 50 μm. Tomoyuki Sumi et al. Development 2008;135: © 2008.
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