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Figure 7. Expression profile of Nodal, Alk4, and ActRIIB during cardiomyocyte differentiation and their effects on cardiac induction. (A) RNA expression levels of Nodal, Alk4, and ActRIIB genes during in vitro differentiation of ES cells. RT-PCR analysis was performed on RNA extracted from either undifferentiated ES or EBs (either wt or Cripto−/−) throughout a differentiation period of 10 d (days 2–10). HPRT gene was used as an internal control. (B) Western blot analysis of total lysates from 293EBNA cells transfected with either wt or ca form of HA-tagged human Alk4. Cells were cotransfected with Jun-HA expression vector as an internal control. A monoclonal anti-HA antibody was used to detect protein levels. (C) RNA expression profile of the αMHC gene during differentiation of Cripto−/− ES cells (days 5, 7, and 10) overexpressing wt and activated forms of either Alk4 or Taram-A. HPRT gene was used as an internal control. Image published in: Parisi S et al. (2003) Copyright © 2003, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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