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. Author manuscript; available in PMC: 2016 Dec 21.
Published in final edited form as: Nature. 2012 Aug 30;488(7413):652–655. doi: 10.1038/nature11333

Figure 2. Parp1 activities during iPSC reprogramming.

Figure 2

a, Functional analysis of Parp1 mutants for rescue of iPSC colony formation in Parp1−/− OSKM-MEFs. Cultures were transduced with green fluorescent protein (GFP), WT Parp1, or Parp1 mutants encoding a catalytic domain missense mutation (CAT mutant; E988K), deletion of the catalytic domain (ΔCAT), deletion of the DNA-binding and automodification domains (CAT-only), or triple missense mutation of the DNA-binding domain (DBD mutant; C21G/C125G/L139P). Zn, zinc fingers; BRCT, BRCA1 carboxy terminus. b, Schematic representation of the Nanog locus transcription start site (TSS) region. Indicated are HpaII/MspI sites (green bars) and amplicons for ‘exon 1/intron 1’ and ‘intron 1’ regions(thick greylines). bp,base pairs. c, Parp1 ChIP analyses of the cultures as indicated, presented as the relative enrichment to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). d, Content of 5hmC assessed by GlucMS-qPCR (as a percentage of total cytosine). e, Content of 5mC, quantified by subtraction of 5hmC content (as in d) from the total methylated cytosine (5mC + 5hmC, as determined by HpaII digestion insensitivity; see Supplementary Fig. 3f). Results in a and c–e are shown as means and s.d. for three independent experiments. Asterisk, P < 0.05; two asterisks, P < 0.01; three asterisks, P < 0.001.