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Fig. 1. Temporal dynamics of the primitive streak and mesoderm progenitors
in β-catenin-activated hES cells. (A) The
Nβ-cateninER cells were cultured in defined medium with or without
4OHT (days 0-3), and the representative morphology at the indicated time
periods was shown. Upper or lower panels shows low or high magnification
images, respectively. Scale bars: 100 µm. (B) Immunostaining
analysis of the day-3 Nβ-cateninER-activated cells. Cells were
cultured with or without 4OHT for 3 days, and subjected to immunostaining with
indicated antibodies, and nuclei were counterstained with diaminopimelic acid
(DAPI). Scale bar: 100 µm. (C) The Nβ-cateninER cells
were cultured with (days 1-3) or without (day 0) 4OHT for the indicated time
periods, and subjected to immunostaining with anti-E-cadherin and
anti-N-cadherin antibodies. Nuclei were counterstained with DAPI. Scale bar:
100 µm. (D) Quantitative real-time PCR (qPCR) analysis on RNA
isolated from undifferentiated Nβ-cateninER cells (day 0) and
β-catenin-activated cells harvested at daily intervals (days 1-5) was
performed using specific primers for the genes indicated. (E)
Immunoblot analysis of cell lysates from Nβ-cateninER cells. Cells
were cultured for the indicated time periods with 4OHT, and cell lysates were
subjected to SDS-PAGE and probed with specific antibodies as indicated.
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