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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 4. — PARP1-HPF1 stimulates histone H3 ADP-ribosylation in LIG1-depleted extracts. (A) Mock-, xLIG1- and xLIG1/xPARP1-depleted extracts were analyzed by immunoblotting using the indicated antibodies. (B) The extracts from (A) were used to replicate sperm nuclei. Chromatin-bound proteins were analyzed by immunoblotting (n = 2). (C) Mock-, xLIG1- and xLIG1/xHPF1-depleted extracts were analyzed by immunoblotting using the indicated antibodies. The asterisk indicates a non-specific band. (D) The extracts from (C) were used to replicate sperm nuclei. Chromatin-bound proteins were analyzed (n = 2). (E) xLIG1/xHPF1-depleted extracts were supplemented with buffer, recombinant His10-xHPF1 (WT), or recombinant mutant His10-xHPF1 (Y251A/R252A) defective for PARP1 binding activity. (F) The extracts from (E) were used to replicate sperm nuclei. Chromatin-bound proteins were analyzed (n = 2).