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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 3. — Reduced pluripotency and enhanced differentiation potential of Parp1^–^/– and Parp7^–/– ES cells. (A) Quantification of ES cells stained for Cdx2, Oct4, Nanog and Stella, as shown in Supplementary Figure S1. Cells were classified as positive or negative and compared using a Chi-squared test with the Yates correction. Cell numbers analysed were Cdx2: n = 8544, 5894 and 5415 for wt (J1 and E14), Parp1^–^/– and Parp7^–^/–, respectively; Oct4: n = 2034, 2978 and 715; Nanog: n = 1210, 2108 and 1032; Stella: n = 1385, 1555 and 1278; P < 0.001 in all cases. (B) Immunofluorescence staining and quantification for Nanog and Oct4 in wt ES cells cultured for 4 days in ES conditions in either the presence or the absence of the poly-ADP-ribose polymerase inhibitor PJ34. A very similar reduction in pluripotency factor expression is observed. Cell numbers were: Nanog: n = 2476 and 2477; P < 0.001; Oct4: n = 2133 and 1992; P = 0.42; Nanog/Oct4 double staining: n = 1718 and 1045; P < 0.001. Scale bars: 20 μm. (C) and (D) RT-qPCR analysis of marker gene expression levels in embryoid bodies generated by hanging drop culture. Note the skewed differentiation trajectories in both Parp1^–^/– and Parp7^–^/– ES cells compared to their background-matched wt control ES cell lines. Parp1-deficient embryoid bodies exhibited a greater variability in size and displayed an overall enhanced speed and/or extent of differentiation towards all embryonic lineages. Parp7 deletion abrogated differentiation towards mesoderm and definitive endoderm (Foxa2, Gata4, Gata6), but overall enhanced ectoderm differentiation.