Fig. 1. Rec8-cohesin is required for crossover-specific dHJ resolution.

a, Experimental strategy. Top row: homolog synapsis with chromatin in blue, green homolog axes, and pink SC central region. The X in the final cartoon indicates a crossover formed after Ndt80 is expressed. Middle row: cell synchronization at the dHJ resolution transition using the inducible NDT80-IN allele; and conditional degradation of target proteins using the AID system. Bottom row: DNA events of meiotic recombination. Only the two chromosomes engaged in recombination are shown.

b, Western analysis of Rec8-AID from subcultures with or without the addition of auxin at 7 hours. Arp7 is used as a loading control.

c, Quantification of Rec8-AID level from the experiment shown in panel b.

d, Quantification of nuclear divisions (MI±MII, cells with two and four nuclei) from REC8-AID subcultures with or without auxin.

e, Representative 1D-gel Southern analysis of crossover (upper panel) and non-crossover (lower panel) formation at the HIS4::LEU2 recombination hotspot from REC8-AID subcultures with or without auxin. ^† cross-hybridizing background bands.

f, Quantification of crossovers and non-crossovers levels from the experiments shown in panel e.

g, Quantification of final levels of crossovers and non-crossovers at 11 hrs from REC8-AID subcultures with or without auxin (mean ± SD, 3 independent experiments, ****P<0.0001, **P=0.0038. two-tailed unpaired t-test).

h, Representative 2D gel Southern analysis of joint molecules from REC8-AID subcultures with or without auxin, before and after release from arrest at 7 and 11 hrs, respectively. The upper left panel highlights joint-molecule species: SEI, single end invasion; IH-dHJ, inter-homolog double Holliday junction; IS-JMs, inter-sister joint molecules; mc-JMs, 3- and 4-chromatid joint molecules.

i, Quantification of total joint molecule levels from experiments represented in panel h (mean ± SD, n=3 independent experiments, NS, not significant, P=0.4561, two-tailed unpaired t-test).