Extended Data Fig. 8: dHJ-ZMM protein interplay suppresses DSB formation.

a, Experimental setup to measure DSB formation by Southern blotting after Yen1^ON expression in ndt80∆ cells. Sae2^AID is depleted by 5-Ph-IAA addition (or DMSO as control) alongside Yen1^ON induction by β-oestradiol addition (or MeOH as control) to prevent endonucleolytic removal of Spo11 and subsequent DSB processing and repair. b, Western blot analysis of Yen1^ON, Sae2^AID and Hop1 protein levels in cells at indicated times in SPM, treated as described in (a). Hop1^P indicates phosphorylated Hop1. Crm1 served as protein loading control. Representative of two biological replicates. c, d, Southern blot analysis of DSB formation at the CCT6 (c) and ERG1 (d) hotspots from (b). Two biological replicates are shown side-by-side in the same blot for each hotspot; replicate 2 of the CCT6 blot corresponds to the cropped blot in Fig. 4d. e, f, Southern blot analysis of DSB formation at the CCT6 (e) and ERG1 (f) hotspots in cells with the indicated genotype, with Msh4^AID depletion by 5-Ph-IAA addition (or DMSO as control) at 7 h in SPM. Two biological replicates are shown per locus; replicate 1 of the CCT6 blot in (e) corresponds to the cropped blot in Fig. 4g. g, Western blot analysis of Zip3^AID, Hop1 and Hop1-pT318 protein levels in cells at indicated times in SPM. Zip3^AID depletion was induced by 5-Ph-IAA addition (or DMSO as control) at 7 h in SPM. Hop1^P indicates phosphorylated Hop1. Pgk1 served as protein loading control. h, As in (g), for ZIP4^AID.