Extended Data Fig. 9: Premature resolution of recombination intermediates disrupts the SC and causes meiosis I failure.

a, Representative FACS analysis of DNA content in cells with the indicated genotype at regular time intervals after induction of meiosis in SPM. Corresponds to the experiment in Fig. 4i–l and Extended Data Fig. 9b–l. b, Time-lapse image montage of Zip1^GFP in a cell nucleus induced to enter meiosis in SPM, with imaging starting at 4.5 h in SPM (t = −30 min). The durations of the zygotene and pachytene stages of prophase I, and the time from pachytene exit to anaphase I are indicated (mean ± s.d of two biological replicates; n = 20 cells each). The brightness of Nup84^mCherry was adjusted to facilitate visibility of the signal. Asterisk indicates the gametogenesis uninherited nuclear compartment (GUNC). Ana, Anaphase. Corresponds to Supplementary Video 8. Scale bar = 1 µm. c, d, Western blot analysis (c) and quantification (d) of Yen1^ON protein levels, normalized to the highest value, in cells at indicated times after Yen1^ON induction by β-oestradiol addition (or MeOH as control) at 7 h in SPM. Error bars represent range of two biological replicates. e, Time-lapse image montage of Zip1^GFP in a cell nucleus after MeOH addition as a control in early to mid-zygotene (t = 0 min, ~5 h in SPM). The brightness of Nup84^mCherry was adjusted to facilitate visibility of the signal. Corresponds to Supplementary Video 10. Scale bar = 1 µm. f, Quantification of structured Zip1^GFP signal from (e). Mean of two biological replicates is plotted (n = 20 cells each). g-i, Quantification of structured Zip1^GFP signal (g), meiosis I entry (h), and sporulation (i) in pachytene cells at the time of β-oestradiol addition (t = 0 min, ~5 h in SPM). Mean of two biological replicates is plotted (n = 20 cells each; error bars represent range). Corresponds to Supplementary Video 11. j-l, Quantification of structured Zip1^GFP signal (j), meiosis I entry (k), and sporulation (l) in pre-leptotene/leptotene cells at the time of β-oestradiol addition (t = 0 min, ~5 h in SPM). Mean of two biological replicates is plotted (n = 20 cells each; error bars represent range). Corresponds to Supplementary Video 12. m, Time-lapse image montage of Zip1^GFP and Htb1^mCherry in a cell nucleus of the indicated genotype. H₂O was added as a control at 7 h in SPM (t = −65 min). Ndt80 expression was induced by β-oestradiol addition at ~8 h in SPM (t = 0 min). Corresponds to Supplementary Video 13. Scale bar = 1 µm. n, Quantification of structured Zip1^GFP signal and the fraction of cells undergoing meiosis I and II based on Htb1^mCherry labelled chromatin from (m) (n = 60 cells). o, As in (m), with CuSO₄ addition to induce expression of Yen1^ON at 7 h in SPM (t = −65 min). Corresponds to Supplementary Video 14. p, As in (n), for (o). q, Quantification of cells with completed or defective meiotic divisions from (n) and (p).