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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Version of Record online: 27 NOV 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 12, pages 2672–2682, December 2012
How to Cite
Ougland, R., Lando, D., Jonson, I., Dahl, J. A., Moen, M. N., Nordstrand, L. M., Rognes, T., Lee, J. T., Klungland, A., Kouzarides, T. and Larsen, E. (2012), ALKBH1 is a Histone H2A Dioxygenase Involved in Neural Differentiation. STEM CELLS, 30: 2672–2682. doi: 10.1002/stem.1228
Author contributions: R.O and E. L.: Conceived and designed the project, performed most of the experiments, wrote the manuscript; D.L.: performed in vitro demethylase activity assay; I. J.: performed apoptosis and differentiation assay; J. A. D and T. R.: performed ChIP-Seq and data analysis; M.N.M.: contributed to data acquisition; L.M.N.: provided mouse lines; E. L., A.K., T.K., and J.T.L.: supervised the project. D.L. and I. J. 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 7, 2012. available online without subscription through the open access option.
- Issue online: 27 NOV 2012
- Version of Record online: 27 NOV 2012
- Accepted manuscript online: 7 SEP 2012 01:07PM EST
- Manuscript Accepted: 19 AUG 2012
- Manuscript Received: 11 APR 2012
- University of Oslo and Norwegian Cancer Society
Additional Supporting Information may be found in the online version of this article.
|sc-12-0349_sm_SupplFigure1.tif||98K||Figure S1: A) Sequences of the primers used in (B) (B) PCR on genomic DNA purified from wild-type and Alkbh1−/− mESCs confirmed the removal of the Alkbh1 gene. Expected size of fragments are 421 base pairs for WT and 897 base pairs for Alkbh1−/− (C) qRT-PCR using an Alkbh1 specific TaqManR probe (Applied Biosystems, assay ID: Mm01296827_m1) showed no presence of the transcript in Alkbh1−/− mESCs.|
|sc-12-0349_sm_SupplFigure2.tif||43K||Figure S2: Differentiation of Alkbh1−/− mESCs leads to decreased viability. Upon induction of differentiation by retinoic acid, the culture of Alkbh1−/− mESCs showed reduced viability as compared to the WT mESCs from differentiation day 2.|
|sc-12-0349_sm_SupplFigure3.tif||38K||Figure S3: ChIP-qPCR validation showing enrichment of four selected ALKBH1 binding regions.|
|sc-12-0349_sm_SupplFigure4.tif||146K||Figure S4: (A) Representative western blots verify the presence of histone methyl modifications after acid extraction. (+ represents 0.5 μg calf thymus histones). The histones were purified from MEF cells cultured under standard conditions in DMEM with 10 % fetal bovine serum. (B) Dot blot immunobinding assay suggest a possible interaction between human ALKBH1 and histone H2A. 0.5 μg of histones were spotted onto nitrocellulose membranes followed by incubation with human ALKBH1. After washing, the remaining ALKBH1 were visualized using an anti-ALKBH1 antibody (Sigma cat. no. A8103)|
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