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. 2020 Sep 25;11:4869. doi: 10.1038/s41467-020-18693-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a Left: whole-mount germlines of the indicated genotypes stained with anti-PAR antibodies and DAPI. Right: PAR staining. Scale bar 20 μm. Analysis was performed in biological duplicates. b Western blot with fractionated extracts shows higher PARG-1^CD::GFP and PARG-1^CD abundance compared to controls. Anti-histone H3 was used as loading control for chromatin-bound fraction. Western blot (bottom) confirmed accumulation of PAR in both parg-1(cd) and parg-1(gk120) null mutants. Analysis was performed in biological duplicates. c Non-deconvolved images of late pachytene nuclei showing SYP-1 and elevated levels of PARG-1^CD::GFP versus PARG-1::GFP with delayed redistribution in late pachytene nuclei. Scale bar 10 μm. Analysis was performed in biological duplicates. d The X-chromosome undergoes desynapsis in parg-1(cd); him-5 double mutants. Arrows indicate the unsynapsed X chromosome in the indicated genotypes before irradiation. Scale bar 10 μm. Analysis was performed in biological duplicates. e Exposure to IR fully rescues X-chromosome desynapsis in parg-1(cd); him-5 double mutants. Scale bar 10 μm. Analysis was performed in biological duplicates. f Quantification of synapsis by SYP-1 and HTP-3 co-staining in the indicated genotypes before and after IR exposure. Error bars indicated S.E.M. Statistical significance was assessed by χ² test (significance level for p < 0.05, ***p < 0.0001, ns = not significant). Number of nuclei analysed (0 Gy-10 Gy): WT (215–205), parg-1(cd) (137–146), him-5 (169–190), parg-1(cd); him-5 (165–212). g Blocking PARG-1 catalytic activity caused milder synergistic effects when combined with him-5 mutants in contrast to parg-1(gk120); him-5. Center of error bars indicates mean. Error bars indicate standard deviation and asterisks show statistical significance calculated by two-tailed nonparametric Mann–Whitney test. (***p < 0.0001). Number of animals scored: parg-1::GFP (5), parg-1(gk120) (5), parg-1(cd)::GFP (4), him-5 (5), parg-1::GFP; him-5 (5), parg-1(gk120); him-5 (9), parg-1(cd); him-5 (10), parg-1(cd)::GFP; him-5 (10). h DAPI-staining and quantification of DAPI-bodies (i) in diakinesis nuclei for the indicated genotypes. Center of error bars indicates mean. Error bars indicate standard deviation and asterisks show statistical significance calculated by two-tailed nonparametric Mann–Whitney test. (***p < 0.0001). Scale bar 5 μm. Number of nuclei analysed is reported in Supplementary Table 2.