Extended Data Fig. 10: Cdc5 promotes SC disassembly in part through the activation of HJ resolvases.
From: Holliday junction–ZMM protein feedback enables meiotic crossover assurance
a, Quantification of Zip1 synapsis and polycomplexes in the absence of Cdc5 induction (MeOH-treated control) at 7 h in SPM (n = 50 nuclei per time point). Controls for the experiments shown in Fig. 5a,b. b, Western blot analysis of Cdc5 and Zip1 protein levels in cells of indicated genotypes at specified times in SPM. Cdc5 was induced by β-oestradiol addition (or MeOH as control) at 7 h in SPM. Pgk1 served as protein loading control. Correspond to experiments in (a) and Fig. 5a,b. c, Quantification of Zip1 protein levels from (b), normalized to the highest value. d, Representative images of meiotic chromosome spreads at indicated times in SPM, immunostained for Zip1 (green). Cdc5 was induced by β-oestradiol addition (or MeOH as control) at 7 h in SPM. Scale bars = 2 µm. e, Quantification of Zip1 synapsis and polycomplexes from (d) (n = 50 nuclei per time point). f, As in (b), for cells of the indicated genotype. Corresponds to experiment in (d) and (e). g, As in (c), for (f). h, i, Time-lapse image montage of Zip1GFP in cell nuclei of indicated genotypes (h) and quantification of structured Zip1GFP signal (i) after Cdc5 induction by β-oestradiol addition at ~7 h in SPM (t = 0 min) (n = 50 cells per genotype). The bottom panel corresponds to Supplementary Video 17. Scale bars = 1 µm. j, Quantification of time to complete loss of structured Zip1GFP signal in cells of indicated genotypes from (i) (mean ± s.d; n = 50 cells per genotype; two-tailed, unpaired Mann-Whitney U test). k, (Top) Representative images of meiotic chromosome spreads of the indicated genotypes at 6 h in SPM, immunostained for Zip1 (green), Rec8 (magenta) and the SPB component γ-tubulin/Tub4 (cyan). Insets show magnified, slightly separated SPBs. Scale bars = 2 µm. (Bottom) Quantification of nuclei with partial or full SPB separation containing Zip1 stretches (weighted mean ± weighted SD of two biological replicates; n = 152–174 nuclei per condition).
