Additional Supporting Information may be found in the online version of this article.

sc-12-0349_sm_SupplFigure1.tif98KFigure 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.tif43KFigure 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.tif38KFigure S3: ChIP-qPCR validation showing enrichment of four selected ALKBH1 binding regions.
sc-12-0349_sm_SupplFigure4.tif146KFigure 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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