Related Experiment Video
Updated: Apr 19, 2026

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
Published on: May 30, 2012
Transient Downregulation of Nanog and Oct4 Induced by DETA/NO Exposure in Mouse Embryonic Stem Cells Leads to
Sergio Mora-Castilla1, Juan R Tejedo2, Rafael Tapia-Limonchi2
1Andalusian Center for Molecular Biology and Regenerative Medicine (CABIMER), University Pablo de Olavide, Biomedical Research Network (CIBER) of Diabetes and Related Metabolic Diseases, Red-Tercel, Avenida Américo Vespucio S/N, 41092 Seville, Spain ; Sanford Consortium for Regenerative Medicine, University of California San Diego, 2880 Torrey Pines Scenic Drive, La Jolla, CA 92037, USA.
Abstract:
The function of pluripotency genes in differentiation is a matter of investigation. We report here that Nanog and Oct4 are reexpressed in two mouse embryonic stem cell (mESC) lines following exposure to the differentiating agent DETA/NO. Both cell lines express a battery of both endoderm and mesoderm markers following induction of differentiation with DETA/NO-based protocols. Confocal analysis of cells undergoing directed differentiation shows that the majority of cells expressing Nanog express also endoderm genes such as Gata4 and FoxA2 (75.4% and 96.2%, resp.). Simultaneously, mRNA of mesodermal markers Flk1 and Mef2c are also regulated by the treatment. Acetylated histone H3 occupancy at the promoter of Nanog is involved in the process of reexpression. Furthermore, Nanog binding to the promoter of Brachyury leads to repression of this gene, thus disrupting mesendoderm transition.
Insights
Pluripotency genes Nanog and Oct4 reexpressed in mouse embryonic stem cells during differentiation. Nanog reexpression influences endoderm marker expression and disrupts mesendoderm transition.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Epigenetics
Background:
- Pluripotency genes maintain embryonic stem cell identity.
- Their role during differentiation is not fully understood.
- Investigating pluripotency gene regulation offers insights into developmental processes.
Purpose of the Study:
- To investigate the role of Nanog and Oct4 in mouse embryonic stem cell (mESC) differentiation.
- To explore the regulatory mechanisms behind Nanog reexpression.
- To understand how Nanog affects lineage specification during differentiation.
Main Methods:
- Used two mouse embryonic stem cell (mESC) lines.
- Induced differentiation using DETA/NO.
- Performed confocal analysis to examine gene expression.
- Assessed histone modifications at gene promoters.
Main Results:
- Nanog and Oct4 were reexpressed in mESCs upon DETA/NO treatment.
- Cells expressed both endoderm (Gata4, FoxA2) and mesoderm (Flk1, Mef2c) markers.
- Nanog reexpression correlated with endoderm marker expression.
- Acetylated histone H3 at the Nanog promoter was involved in its reexpression.
- Nanog repressed Brachyury, disrupting mesendoderm transition.
Conclusions:
- Nanog and Oct4 reexpression occurs during mESC differentiation.
- Nanog plays a role in endoderm specification and mesendoderm transition.
- Histone acetylation is implicated in Nanog reexpression.
- This study reveals a complex regulatory role for pluripotency factors during differentiation.
More Related Videos
Related Concept Videos
Forced Transdifferentiation
Artificial...
Maintenance of the ES Cell State

