(a) Purified core histones from rad53-AID and rad53-AID lsm1∆ cells that were treated or not 2 hr with NAA and blocked with Nocodazole 2 additional hours. The image represents a Coomassie-Blue-stained gel of histone-purified samples. (b) Quantification of histones in the samples loaded in (a). Histone signal (indicated with a bracket) was normalised using the signal of a non-specific band (indicated with an asterisk). The results were normalised to lane 1. Quantification was done using image J. Two independent experiments were performed. *p=0.026 (t-test, paired, two tails) (c) Western Blots of canonical histones H3, H4 and H2A, and the histone variant H2A.Z. performed with purified core histones samples. Ratio H2A/H4 and H2AZ/H4 are shown in Figure 3—figure supplement 3f. (d) Micrococcal nuclease (Mnase) analysis of chromatin extracted from rad53-AID and rad53-AID lsm1∆ cells treated as described in (a). (e) Live microscopy (20-min interval, 14 Z-sections, 2,89µ total) in strain DVY36 (rad53-AID lsm1∆ NUP49-GFP) after 90 min of Auxin treatment (Auxin was maintained in the media). (f) Cell cycle distribution of rad53-AID, lsm1∆ and rad53-AID lsm1∆ cells after treatment with 500 µM NAA. (g) Pulse field gel electrophoresis of rad53-AID lsm1∆ cells that were treated with 200 mM Hydroxyurea (HU), 10 µg/ml Nocodazole (NOC), or 500 µM NAA (NAA). Control asynchronous growing cells were not treated (UT). HU-treated cells are blocked in S-phase preventing chromosomes from entering the gel. Nocodazole allows full replication but blocks mitosis. (h) Percentage of rad53-AID lsm1∆ cells that have experienced or not a WGD after a 4 hr treatment with 500 µM NAA. The DNA content of all survivors was analysed by FACS. two independent experiments (n = 100 or 75) p-value=0.038 (t-test, two tails, unequal variance).