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. 2021 Apr 2;220(6):e202012057. doi: 10.1083/jcb.202012057

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© 2021 Dello Stritto et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 6. — Impairment of the NHEJ/Alt-NHEJ/SSA pathways in meiosis triggers p53/CEP-1-dependent apoptosis. (A) Apoptosis quantification using SYTO-12 in the indicated genotypes. n = number of gonads scored. top-3(jf101),4.2 ± 1.9, n = 43 (quantification as in Fig. 2 D); cku-80(ok861),2.9 ± 1.3, n = 65; cku-80(ok861) top-3(jf101),8.4 ± 3, n = 47; cku-70(tm1524),2.2 ± 1.3, n = 24; cku-70(tm1524) top-3(jf101),8.2 ± 2.6, n = 52; lig-4(ok716),2.2 ± 1, n = 52; top-3(jf101) lig-4(ok716),7.2 ± 2.6, n = 98; polq-1(tm2026),2.3 ± 1.1, n = 65; polq-1(tm2026) top-3(jf101),6 ± 2.1, n = 70; cku-70(tm1524) polq-1(tm2026),3.2 ± 1.6, n = 80; and top-3(jf101) cku-70(tm1524) polq-1(tm2026),7.3 ± 2.8, n = 83; xpf-1(tm2842), 4.1 ± 1.7, n = 52; top-3(jf101); xpf-1(tm2842), 6.7 ± 3.4, n = 56. ****, P < 0.0001; ns,calculated using the Mann–Whitney test. (B) Apoptosis quantification with SYTO-12 in the indicated genotypes (quantification as described for Fig. 4 C for wt, wt; spo-11^RNAi; top-3(jf101) and top-3(jf101); spo-11^RNAi): n = number of gonads scored: wt, 4.5 ± 2.3, n = 41; wt; spo-11^RNAi, 4.5 ± 2.3, n = 78; top-3(jf101), 5.3 ± 1.9, n = 20; top-3(jf101); spo-11^RNAi, 5.2 ± 2.6, n = 89; top-3(jf101) cku-80(ok861),9.2 ± 2.5, n = 18; and top-3(jf101) cku-80(ok861); spo-11^RNAi, 4.8 ± 2, n = 88. ****, P < 0.0001; ns, calculated using the Mann–Whitney test. Error bars indicate mean ± SD. (C) Apoptosis quantification with SYTO-12 in the indicated genotypes. n = number of gonads scored: cep-1(gk138),2.1 ± 1.2, n = 67; top-3(jf101), 4.2 ± 2, n = 43; cep-1(gk138); top-3(jf101), 1.9 ± 1, n = 66; cku-80(ok861) top-3(jf101), 8.5 ± 3.5, n = 25; and cep-1(gk138); top-3(jf101) cku-80(ok861),1.9 ± 1.2, n = 58. Error bars indicate the mean (and SD), ****, P < 0.0001 calculated using the Mann–Whitney test. (D) Model showing the function of topoisomerase 3 during mitosis and meiosis (green to blue gradient). In mitosis, TOP-3 is involved in processing stalled and collapsed replication forks; in meiosis, it interferes with strand invasion and is involved in dissolving joint molecules. In mitosis, top-3 mutants accumulate aberrant recombination intermediates that are probably coated with RAD-51. Those abnormal intermediates are imported into meiosis, where they are mostly refractory to apoptosis. In meiosis, HIM-6 generates aberrant intermediates that are directed toward repair via the NHEJ, Alt-NHEJ, and SSA pathways in the top-3 mutant, which prevents efficient apoptosis. wt, wild-type.