Correlations Between the Levels of Oct4 and Nanog as a Signature for Naïve Pluripotency in Mouse Embryonic Stem Cells§


  • Author contributions: S.M.D.: conceived the project, carried out the experiments, the measurements, and the analysis of the data, and wrote the manuscript; A.M.A.: conceived the project and wrote the manuscript; P.R. and J.G.O.: contributed to the analysis of the data.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS November 7, 2012.


The pluripotent state is traditionally associated with large absolute levels of certain transcription factors such as Nanog and Oct4. Here, we present experimental observations using quantitative immunofluorescence that pluripotency in mouse embryonic stem cells (mESCs) is established by specific ratios between Oct4 and Nanog. When cells are grown in 2i conditions, they exhibit uniform levels of pluripotency and this is associated with a high correlation between the levels of Oct4 and Nanog in individual cells. The correlation is lost when cells differentiate. Our results suggest that the correlation between these two factors and the distribution of Oct4/Nanog ratios can be used as quantifiers to distinguish between three subpopulations in an mESC culture: pluripotent, lineage-primed, and differentiating cells. When we apply these quantifiers to cells with lower levels of Nanog or mutant for β-Catenin or Tcf3, the results suggest that these cells exhibit higher probability of differentiation. STEM CELLS 2012;30:2683–2691