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. 2014 Jul 17;42(14):8914–8927. doi: 10.1093/nar/gku591

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Identification of Parp1 and Parp7 as ES cell-associated genes. (A) RT-qPCR analysis of Parp family members in ES and TS cells. Data of 3–6 independent biological replicates are represented as mean + SEM (**P < 0.01). (B) Northern blot analysis of Parp7 expression in two different ES (B6 and RRG177) and TS (Rs26 and GFP) cell lines. (C) RT-qPCR analysis of Parp1 and Parp7 expression in wildtype (J1 and E14) ES cells over a differentiation time course of 8 days induced by LIF withdrawal and culture on non-adherent plates. (D) Schematic representation of the Parp1 and Parp7 protein domain structure. PADR1 = PADR1 domain; BRCT = BRCA1 C-Terminus domain; WGR = WGR domain; WWE = WWE domain. (E) Immunolocalization of Parp1 and Parp7, detected by confocal microscopy after staining with antibodies against the C-terminal FLAG-tag as well as against the endogenous proteins, showing the nuclear localisation of Parp1 and Parp7. Note that the antibody against endogenous Parp7 is not particularly efficient. Scale bar: 25 μm. (F) Genotyping PCR proving establishment of a homozygously gene-trapped ES cell line at the Parp7 locus. (G) RT-qPCR expression levels of Parp7 in wildtype (wt), Parp7^+/− and Parp7^-^/– ES cells (*P < 0.05, ***P < 0.001).