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| Status |
Public on Feb 07, 2011 |
| Title |
MeDIP-on-chip and Pol-II ChIP-on-chip during C2C12 myogenic and BMP2-induced osteogenic differentiation |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
Methylation profiling by genome tiling array
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| Summary |
Differentiation of pluripotent embryonic stem cells, via more restricted multipotent adult stem cells, towards the multitude of terminally differentiated, specialized cell types that make up the adult body is a multi-step process characterized by a progressive restriction of differentiation potential. Previous studies have demonstrated tissue-specific differences in DNA methylation patterns that might play a role in lineage restriction and tissue-specific gene expression. However, these studies have mainly focused on either the differentiation of pluripotent ES cells or on comparing somatic cells from different tissues, so that it remains unclear at which stages during development these differences in DNA methylation profiles are established. Since data on the role of DNA methylation during late-stage development is limited, we have examined the role of this epigenetic mark during the terminal differentiation of a multipotent adult stem cell, using C2C12 as a model system. Using parallel MeDIP- and Pol-II ChIP-on-chip approaches, we demonstrate a mutually exclusive relationship between high Pol-II occupancy and high DNA methylation levels. In addition, we observe that in undifferentiated progenitor cells, osteoblast-related genes have in general higher methylation levels than myoblast-related genes, while myoblast-related genes have higher Pol-II occupancy. Induction of osteogenesis with BMP2 leads to a rearrangement of Pol-II towards osteoblast-related genes. However, methylation patterns remain unchanged. These results indicate that DNA methylation primes C2C12 multipotent stem cells for myogenic differentiation and permits induction of the osteogenic gene expression program upon growth factor stimulation. We propose that cell type-specific DNA methylation patterns are established prior to terminal differentiation of adult stem cells.
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| Overall design |
C2C12 cells were grown for 0, 1, 3 and 6 days in the presence or absence of BMP2. DNA was isolated and ChIPed with anti-Pol-II or IP-ed with anti-5-methyl-cytosine. non-IP'd input DNA was used as a reference sample
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| Contributor(s) |
Hupkes M, van Someren EP, Piek E, Dechering KJ, van Zoelen EJ |
| Citation(s) |
21277338 |
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| Submission date |
Jun 01, 2010 |
| Last update date |
Oct 14, 2014 |
| Contact name |
Marlinda Hupkes |
| E-mail(s) |
[email protected]
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| Phone |
+31(0)243652519
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| Organization name |
Radboud University Nijmegen
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| Department |
Cell and Applied Biology
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| Street address |
Heyendaalseweg 135
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| City |
Nijmegen |
| ZIP/Postal code |
6525 AJ |
| Country |
Netherlands |
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| Platforms (1) |
| GPL5811 |
[Mm_PromPR] Affymetrix Mouse Promoter 1.0R Array |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA128959 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE22077_ChIP_Tx_mm9.bed.gz |
2.9 Mb |
(ftp)(http) |
BED |
| GSE22077_Me_mm9.bed.gz |
1.5 Mb |
(ftp)(http) |
BED |
| GSE22077_RAW.tar |
787.6 Mb |
(http)(custom) |
TAR (of BAR, CEL) |
| Processed data provided as supplementary file |
| Processed data are available on Series record |
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