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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Article first published online: 12 FEB 2013
Copyright © 2012 AlphaMed Press
Volume 31, Issue 2, pages 282–292, February 2013
How to Cite
Park, K.-S., Cha, Y., Kim, C.-H., Ahn, H.-J., Kim, D., Ko, S., Kim, K.-H., Chang, M.-Y., Ko, J.-H., Noh, Y.-S., Han, Y.-M., Kim, J., Song, J., Kim, J. Y., Tesar, P. J., Lanza, R., Lee, K.-A. and Kim, K.-S. (2013), Transcription Elongation Factor Tcea3 Regulates the Pluripotent Differentiation Potential of Mouse Embryonic Stem Cells Via the Lefty1-Nodal-Smad2 Pathway. STEM CELLS, 31: 282–292. doi: 10.1002/stem.1284
Author contributions: K.-S.P. and K.-S.K.: concept and design, data analysis and interpretation, and writing; Y.C.: concept and design, collection and/or assembly of data, and data analysis and interpretation; C.-H.K., H.-J.A., D.K., S.K., K.-H.K., M.-Y.C., J.-H.K., J.Y.K., J.K., and J.S.: collection and/or assembly of data; Y.-S.N., Y.-M.H., P.J.T., R.L., and K.-A.L.: data analysis and interpretation. K.-S.P. and Y.C. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS November 25, 2012.
- Issue published online: 12 FEB 2013
- Article first published online: 12 FEB 2013
- Accepted manuscript online: 21 NOV 2012 12:33AM EST
- Manuscript Accepted: 25 OCT 2012
- Manuscript Received: 6 JUL 2012
- Korea Science and Engineering Foundation (KOSEF)
- Korean government (MEST). Grant Numbers: 2012-050367, 2012-0005261
- Priority Research Centers Program through the National Research Foundation of Korea (NRF)
- Ministry of Education, Science, and Technology. Grant Number: 2012-0006679
- National Institute of Health (NIH). Grant Numbers: MH087903, HL106627, NS070577
Additional Supporting Information may be found in the online version of this article.
|sc-12-0609_sm_SupplFigure1.tif||1226K||Figure S1. A customized SuperArray membrane was constructed with 111 selected genes based on genome-wide transcriptome analysis of mouse oocytes during in vitro maturation.|
|sc-12-0609_sm_SupplFigure2.tif||551K||Figure S2. mESCs were differentiated into mesoendoderm lineage for 4 days and markers representing mesoderm and endoderm were analyzed for the expression by real-time RT-PCR. All values are means ±;s.d. from at least triplicate experiments. * indicates significant (P<0.05) and ** highly significant (p<0.01) results based on ANOVA analyses following the Scheffe test.|
|sc-12-0609_sm_SupplFigure3.tif||606K||Figure S3. 4 weeks old teratoma of WT and Tcea3 KD were analyzed for the expression of markers representing each lineage by real-time RT-PCR. The expression level of each gene was shown as relative value following normalization against that of the glyceraldehyde 3-phosphate dehydrogenase (Gapdh) gene.|
|sc-12-0609_sm_SupplFigure4.tif||938K||Figure S4. Chromatin immunoprecipitation using antibodies against Tcea3 and rabbit IgG (as an immunoprecipitation control). DNA fragments were analyzed by PCR using primers specific for the proximal promoter region (Chip1) and downstream region of the Lefty1 gene (Chip2) (left). Relative quantification of Tcea3 immunoprecipitation enrichment at the Chip1 and Chip2 region was performed with WT, Tcea3 KD and Tcea3 OE cells. All values are means ±;s.d. from at least triplicate experiments. * indicates significant (P<0.05) results based on Student's T-test analyses.|
|sc-12-0609_sm_SupplFigure5.tif||1796K||Figure S5. (A) Immunoblot analysis of p-Smad1/5 in Tcea3 OE and WT mESCs. Total cell extracts were prepared either from undifferentiated cells or 4 days after RA-induced differentiation. β-actin was used as loading control. (B) Expression levels of BMP4, ID1, ID2 and ID3 by microarray analysis of Tcea3 OE cells. (C) WT and Tcea3 OE cells were serum starved for 24 hrs and stimulated with Nodal (1 μg/ml). Cells were harvested at the indicated times after Nodal stimulation and pSmad2 (left panel) or pSmad1/5 (right panel) were analyzed by immunoblot.|
|sc-12-0609_sm_SupplFigure6.tif||62K||Figure S6. Microarray analysis of Tcea3 OE shows mesoderm marker genes of Tcea3 OE are activated compared to wild type mESCs under self-renewing condition.|
|sc-12-0609_sm_SupplTable1.pdf||55K||Table S1. Selected candidates identified as binding proteins of Tcea3.|
|sc-12-0609_sm_SupplTable2.pdf||60K||Table S2. 359 genes whose expression was significantly up- or down- regulated in Tcea3 OE mESCs.|
|sc-12-0609_sm_SupplTable3.pdf||11K||Table S3. RT-PCR primers used in this study|
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