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FOXC1 Downregulates Nanog Expression by Recruiting HDAC2 to Its Promoter in F9 Cells Treated by Retinoic Acid

Hongni Xue1,2, Fayang Liu1,2, Zhiying Ai1,2

  • 1College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China.

International Journal of Molecular Sciences
|March 6, 2021
PubMed

Insights

Forkhead box C1 (FOXC1) directly inhibits Nanog, a key pluripotency factor, during cell differentiation. FOXC1 recruits HDAC2 to the Nanog promoter, epigenetically silencing gene expression.

Area of Science:

  • Cell Biology
  • Epigenetics
  • Developmental Biology

Background:

  • FOXC1 is a transcription factor crucial for cell differentiation and embryogenesis.
  • Nanog is a core pluripotency factor essential for maintaining embryonic stem cell identity.

Purpose of the Study:

  • To investigate the regulatory relationship between FOXC1 and Nanog.
  • To elucidate the molecular mechanism by which FOXC1 influences Nanog expression.

Main Methods:

  • Analysis of FOXC1 expression during retinoic acid-induced differentiation of F9 Embryonal Carcinoma (EC) cells.
  • Overexpression of FOXC1 in F9 cells and knockdown in 3T3 cells to assess Nanog mRNA and protein levels.
  • Identification of FOXC1 interacting proteins using interactome analysis.
  • Chromatin immunoprecipitation (ChIP) assays to examine histone modifications and protein recruitment to the Nanog promoter.

Main Results:

  • FOXC1 is upregulated during F9 EC cell differentiation.
  • FOXC1 directly binds to the Nanog promoter and inhibits its transcription.
  • Overexpression of FOXC1 downregulates Nanog, while its knockdown upregulates Nanog.
  • HDAC2 was identified as a FOXC1 co-repressor.
  • FOXC1 recruits HDAC2 to the Nanog promoter, leading to decreased H3K27ac enrichment and transcriptional repression.

Conclusions:

  • FOXC1 acts as a negative regulator of Nanog during cell differentiation.
  • FOXC1-mediated repression of Nanog involves the recruitment of the co-repressor HDAC2.
  • This study reveals a novel mechanism of epigenetic regulation by FOXC1 impacting pluripotency factor expression.
Keywords:
ARTAF9 cellsFOXC1 interactomeHDAC2Nanogpromoter analysis

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