Abstract
The first cell fate choice in the mammalian embryo, the segregation of the inner cell mass (ICM) and trophectoderm (TE), is regulated by the mutually antagonistic effects of the transcription factors, Oct4 and Cdx2, while the pluripotency factor, Nanog, is essential to specify the epiblast. We have analyzed the promoters of Nanog and Cdx2, and have found that these two transcription factors are likewise regulated reciprocally. Using an embryonic stem cell line with conditional TE differentiation, we show that Nanog overexpression suppresses the upregulation of TE markers, while Nanog knockdown upregulates the expression of TE markers. We further show that Nanog and Cdx2 bind to and repress each other's promoters. However, whereas Nanog knockout results in detectable Cdx2 expression in the ICM, we observe no overt disruption of blastocyst development, indicating that Nanog plays a subservient role to Oct4 in segregation of the ICM and TE.
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Acknowledgements
GQD was supported by grants from the National Institutes of Health, the NIH Director's Pioneer Award of the NIH Roadmap for Medical Research, and private funds contributed to the Harvard Stem Cell Institute and the Children's Hospital Stem Cell Program, USA. GQD is a recipient of Clinical Scientist Awards in Translational Research from the Burroughs Wellcome Fund and the Leukemia and Lymphoma Society, and is an Investigator at the Howard Hughes Medical Institute. KH was supported by the NIH Director's Innovator Award and the Harvard Stem Cell Institute, USA.
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Supplementary information
Supplementary information, Figure S1
Schematic illustration of the inducible system used in this study. (PDF 90 kb)
Supplementary information, Figure S2
Nanog over-expression quantitatively suppresses the up-regulation of TE markers by Ras or Cdx2. (PDF 141 kb)
Supplementary information, Figure S3
Knockdown of Nanog by Stealth RNAi results in up-regulation of Cdx2, as well as Gata6. (PDF 151 kb)
Supplementary information, Figure S4
Mapping Nanog and Oct4 binding sites in the Cdx2 locus. (PDF 129 kb)
Supplementary information, Figure S5
Immunofluorescence of Nanog−/− embryos. (PDF 120 kb)
Supplementary information, Data S1
Primers for cDNA quantification (PDF 35 kb)
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Chen, L., Yabuuchi, A., Eminli, S. et al. Cross-regulation of the Nanog and Cdx2 promoters. Cell Res 19, 1052–1061 (2009). https://doi.org/10.1038/cr.2009.79
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DOI: https://doi.org/10.1038/cr.2009.79
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