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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 S1. — top-3 mutants display joint DNA structures at diakinesis that are independent of the major and alternative DNA repair pathways. Related to Fig. 1. (A) Representative images of gonads stained with DAPI of the indicated genotypes. *, Mitotic tip. Dashed line marks the mitotic gonad region. (B) FISH analysis of wt and top-3(jf101) diakinesis using two different probes: chromosome III (magenta) and chromosome V (cyan). Dashed circles highlight single DAPI bodies that were assessed in the image stacks before projection. (C and D) Quantification of DAPI-stained bodies in −1 diakinesis oocyte nuclei of different genotypes. Scatter plots indicate the mean ± SD; n = number of oocytes for each genotype. (C) top-3(jf101) (as shown in Fig. 1 C), 5.5 ± 0.9, n = 66; brc-1(tm1145), 6 ± 0, n = 20; brc-1(tm1145) top-3(jf101), 5.9 ± 0.3, n = 104; mus-81(tm1937), 6.3 ± 0.6, n = 21; mus-81(tm1937); top-3(jf101), 4.3 ± 1.02, n = 26; xpf-1(tm2842), 6.5 ± 0.6, n = 30; xpf-1(tm2842); top-3(jf101), 5.6 ± 0.7, n = 21; polq-1(tm2026), 6.06 ± 0.2, n = 35; polq-1(tm2026) top-3(jf101), 6.06 ± 0.6, n = 34; cku-80(ok861), 6 ± 0, n = 26; top-3(jf101) cku-80(ok861), 5.6 ± 0.6, n = 88; cku-70(tm1524), 6 ± 0, n = 38; top-3(jf101) cku-70(tm1524), 5.9 ± 0.4, n = 34; polq-1(tm2026) cku-70(tm1524), 6.02 ± 0.1, n = 65; and top-3(jf101) polq-1(tm2026) cku-70(tm1524) 5.5 ± 0.8, n = 55. (D) zhp-3(jf61), 11.7 ± 0.4, n = 38; zhp-3(jf61); top-3(jf101), 6.5 ± 0.8, n = 35; zhp-3(jf61); brc-1(tm1145) top-3(jf101), 6.8 ± 0.9, n = 45; zhp-3(jf61); top-3(jf101); cku-80^RNAi, 6.7 ± 0.9, n = 32; and zhp-3(jf61); top-3(jf101) brc-1(tm1145); cku-80^RNAi, 6.7 ± 0.8, n = 31. Note that the mean number of DAPI diakinesis bodies is similar for all indicated double and triple mutants and that the configuration of DAPI bodies in diakinesis (undefined DNA masses) resembles those seen in top-3, also in the absence of CO formation. Error bars indicate the mean ± SD per genotype. Comparison calculated using the Mann–Whitney test: top-3(jf101) versus brc-1(tm1145) top-3(jf101), ****, P < 0.0001; top-3(jf101) versus mus-81(tm1937); top-3(jf101), ****, P < 0.0001; top-3(jf101) versus xpf-1(tm2842); top-3(jf101), ns; top-3(jf101) versus polq-1(tm2026) top-3(jf101), ***, P = 0.0001; top-3(jf101) versus top-3(jf101) cku-80(ok861), *, P = 0.027; top-3(jf101) versus top-3(jf101) cku-70(tm1524),***, P = 0.0002; top-3(jf101) versus top-3(jf101) polq-1(tm2026) cku-70(tm1524), ns. Chr, chromosome; wt, wild-type.