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. 2021 Apr 19;49(9):5003–5016. doi: 10.1093/nar/gkab269

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© The Author(s) 2021. 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 5. — HPF1-dependent PARP activation promotes alternative Okazaki fragment ligation. (A) Mock-, xLIG1-, xPARP1- and xLIG1/xPARP1-depleted extracts were used for replication of sperm nuclei. Chromatin-bound proteins were analyzed by immunoblotting using indicated antibodies (n = 2). (B) Mock-, xLIG1-, xHPF1- and xLIG1/xHPF1-depleted extracts were used for replication of sperm nuclei. Chromatin-bound proteins were analyzed by immunoblotting using the indicated antibodies (n = 2). (C) xLIG1-, xLIG1/xXRCC1-, xLIG1/xPARP1- or xLIG1/xHPF1-depleted extracts were used to replicate sperm chromatin. Genomic DNA was purified and labeled using exonuclease-deficient Klenow fragments and α-³²P dCTP. Samples were separated in a denaturing agarose gel, and the incorporation of radioactivity was monitored by autoradiography (n = 3). (D) Replication products in mock-, xLIG1-, xLIG1/xXRCC1-, xPARP1-, xLIG1/xPARP1-, xHPF1- or xLIG1/xHPF1-depleted extracts were separated by alkaline agarose gel electrophoresis followed by autoradiography (n = 4).