J. Ms#-200901040r2, Online Supplemental material S1: Characterization of HUES-9 pOct4/GFP clone. Non-cardiac potential of differentiation of Oct4-OEC

. Sox2, Sox17 and consequences on BMP2-induced gene expression S2: Paracrine action of Oct4-overexpressing cells S3: Oct4 overexpressing cells activate smad and ?-catenin pathways; The Oct4

D. We and H. Ng, Genome Institute of Singapore) for the gift of Sox2 ShRNA plasmid, Drs M Mitjavila and N Lefort for help in FACS analysis, Y Maury (INSERM-UMR861) for help in HC imaging

R. Singapore, . Feil, and . Montpellier, France) for critical reading of the MS, Dr M. Buckingham (Pasteur Institute, Paris) for her invaluable comments on the MS the National Agency for Research (ANR, programmes Blanc, HUES signature and Specistem) and Genopole for funding this research. SS and DN were funded by ANR, p.18

J. Ms#-200901040r2-references-bakre, M. M. , A. Hoi, J. C. Mong, Y. Y. Koh et al., Generation of multipotential mesendodermal progenitors from mouse embryonic stem cells via sustained Wnt pathway activation, J Biol Chem, vol.282, pp.31703-31715, 2007.

E. Ben-shushan, H. Sharir, E. Pikarsky, and Y. Bergman, A dynamic balance between ARP-1/COUP-TFII, EAR-3/COUP-TFI, and retinoic acid receptor:retinoid X receptor heterodimers regulates Oct-3/4 expression in embryonal carcinoma cells., Molecular and Cellular Biology, vol.15, issue.2, pp.1034-1082, 1995.
DOI : 10.1128/MCB.15.2.1034

A. Bondue, G. Lapouge, C. Paulissen, C. Semeraro, M. Iacovino et al., Mesp1 Acts as a Master Regulator of Multipotent Cardiovascular Progenitor Specification, Cell Stem Cell, vol.3, issue.1, pp.69-84, 2008.
DOI : 10.1016/j.stem.2008.06.009

H. H. Robson and . Ng, Reciprocal transcriptional regulation of Pou5f1 and Sox2 via the Oct4/Sox2 complex in embryonic stem cells, Mol Cell Biol, vol.25, pp.6031-6077, 2005.

D. J. Crease, S. Dyson, and J. B. Gurdon, Cooperation between the activin and Wnt pathways in the spatial control of organizer gene expression, Proceedings of the National Academy of Sciences, vol.95, issue.8, pp.4398-403, 1998.
DOI : 10.1073/pnas.95.8.4398

J. A. Dahl and P. Collas, ChIP, a Quick and Quantitative Chromatin Immunoprecipitation Assay, Unravels Epigenetic Dynamics of Developmentally Regulated Genes in Human Carcinoma Cells, Stem Cells, vol.6, issue.4, pp.1037-1083, 2007.
DOI : 10.1634/stemcells.2006-0430

L. M. Hoffman and M. K. Carpenter, Characterization and culture of human embryonic stem cells, Nature Biotechnology, vol.145, issue.6, pp.699-708, 2005.
DOI : 10.1093/hmg/ddh088

M. Kasai, K. Satoh, and T. Akiyama, Wnt signaling regulates the sequential onset of neurogenesis and gliogenesis via induction of BMPs, Genes to Cells, vol.10, issue.8, pp.777-83, 2005.
DOI : 10.1016/S0012-1606(03)00123-4

J. Lee, H. K. Kim, J. Y. Rho, Y. M. Han, and J. Kim, The Human OCT-4 Isoforms Differ in Their Ability to Confer Self-renewal, Journal of Biological Chemistry, vol.281, issue.44, pp.33554-65, 2006.
DOI : 10.1074/jbc.M603937200

Y. Liu, M. Asakura, H. Inoue, T. Nakamura, M. Sano et al., Sox17 is essential for the specification of cardiac mesoderm in embryonic stem cells, Proceedings of the National Academy of Sciences, vol.104, issue.10, pp.3859-64, 2007.
DOI : 10.1073/pnas.0609100104

N. Maherali, T. Ahfeldt, A. Rigamonti, J. Utikal, C. Cowan et al., A High-Efficiency System for the Generation and Study of Human Induced Pluripotent Stem Cells, Cell Stem Cell, vol.3, issue.3, pp.340-345, 2008.
DOI : 10.1016/j.stem.2008.08.003

C. Menard, C. Grey, A. Mery, D. Zeineddine, F. Aimond et al., Cardiac specification of embryonic stem cells, Journal of Cellular Biochemistry, vol.77, issue.IV, pp.681-687, 2004.
DOI : 10.1002/jcb.20259

S. W. Willemse, A. H. De-laat, and . Piersma, Visceral-endoderm-like cell lines induce differentiation of murine P19 embryonal carcinoma cells, Differentiation, vol.46, pp.51-60, 1991.

A. T. Naito, I. Shiojima, H. Akazawa, K. Hidaka, T. Morisaki et al., Developmental stage-specific biphasic roles of Wnt/beta-catenin signaling in cardiomyogenesis and hematopoiesis, Proceedings of the National Academy of Sciences, vol.103, issue.52, pp.19812-19819, 2006.
DOI : 10.1073/pnas.0605768103

J. D. Nelson, O. Denisenko, and K. Bomsztyk, Protocol for the fast chromatin immunoprecipitation (ChIP) method, Nature Protocols, vol.34, issue.1, pp.179-85, 2006.
DOI : 10.1038/nprot.2006.27

H. Niwa, How is pluripotency determined and maintained? Development, pp.635-681, 2007.
DOI : 10.1242/dev.02787

URL : http://dev.biologists.org/cgi/content/short/134/4/635

H. Niwa, J. Miyazaki, and A. G. Smith, Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells, Nature Genetics, vol.24, issue.4, pp.372-378, 2000.
DOI : 10.1038/74199

M. C. Nostro, X. Cheng, G. M. Keller, and P. Gadue, Wnt, Activin, and BMP Signaling Regulate Distinct Stages in the Developmental Pathway from Embryonic Stem Cells to Blood, Cell Stem Cell, vol.2, issue.1, pp.60-71, 2008.
DOI : 10.1016/j.stem.2007.10.011

. Zandstra, Niche-mediated control of human embryonic stem cell self-renewal and differentiation, Embo J, vol.26, pp.4744-55, 2007.

M. W. Pfaffl, A new mathematical model for relative quantification in real-time RT-PCR, Nucleic Acids Research, vol.29, issue.9, p.45, 2001.
DOI : 10.1093/nar/29.9.e45

M. Puceat, S. Roche, and G. Vassort, Src Family Tyrosine Kinase Regulates Intracellular pH in Cardiomyocytes, The Journal of Cell Biology, vol.108, issue.7, pp.1637-1683, 1998.
DOI : 10.1074/jbc.270.41.24428

C. A. Seguin, J. S. Draper, A. Nagy, and J. Rossant, Establishment of Endoderm Progenitors by SOX Transcription Factor Expression in Human Embryonic Stem Cells, Cell Stem Cell, vol.3, issue.2, pp.182-95, 2008.
DOI : 10.1016/j.stem.2008.06.018

A. G. Smith, Culture and differentiation of embryonic stem cells, Journal of Tissue Culture Methods, vol.336, issue.2, pp.89-94, 1991.
DOI : 10.1007/BF01666137

T. Sumi, N. Tsuneyoshi, N. Nakatsuji, and H. Suemori, Defining early lineage specification of human embryonic stem cells by the orchestrated balance of canonical Wnt/??-catenin, Activin/Nodal and BMP signaling, Development, vol.135, issue.17, pp.2969-79, 2008.
DOI : 10.1242/dev.021121

K. Takahashi, K. Tanabe, M. Ohnuki, M. Narita, T. Ichisaka et al., Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors, Cell, vol.131, issue.5, pp.861-72, 2007.
DOI : 10.1016/j.cell.2007.11.019

P. Tam and G. Schoenwolf, Cardiac fate map: lineage, allocation, morphogenetic movement and cell commitment, Heart, vol.Development, pp.3-18, 1999.

J. Hagège, P. Itskovitz-eldor, M. Menasché, and . Pucéat, Differentiation in vivo of cardiac committed Human embryonic stem cells in post-myocardial infarcted rats, Stem Cells, vol.25, pp.2200-2205, 2007.

S. Yamanaka, Strategies and New Developments in the Generation of Patient-Specific Pluripotent Stem Cells, Cell Stem Cell, vol.1, issue.1, pp.39-49, 2007.
DOI : 10.1016/j.stem.2007.05.012

D. Zeineddine, E. Papadimou, K. Chebli, M. Gineste, J. Liu et al., Oct-3/4 Dose Dependently Regulates Specification of Embryonic Stem Cells toward a Cardiac Lineage and Early Heart Development, Developmental Cell, vol.11, issue.4, pp.535-581, 2006.
DOI : 10.1016/j.devcel.2006.07.013

A. M. Zorn, G. D. Barish, B. O. Williams, P. Lavender, M. W. Klymkowsky et al., Regulation of Wnt Signaling by Sox Proteins, Molecular Cell, vol.4, issue.4, pp.487-98, 1999.
DOI : 10.1016/S1097-2765(00)80200-2