Supplementary Figure 4 : Cohesin-dependent compaction is independent of sister chromatid cohesion.

From: SMC complexes differentially compact mitotic chromosomes according to genomic context

Supplementary Figure 4

(a) FACS for DNA content (left) and Western blotting (right) showing that cdc45 and cd45 scc1-73 cells enter mitosis without DNA replication, with CDK phosphorylating condensin on Smc4 Serine 4 (Smc4 S4P) with the same kinetics as wildtype cells ( unspecific band). Picture of Ponceau stained blot confirms equal loading of Western (right, bottom). Western blotting for to confirm CDK activation in cells was from one experiment. (b) Plot of nuclear morphology examining number of DAPI stained cells from the indicated timepoints that have undergone nuclear division. % of single nucleus (rectangles), double nuclei (checked line with squares) and cells with an anaphase nucleus are shown (grey line). (c) (Top) Zoom into contact heatmaps from cdc45 mitotically arrested cells, C, (top) and cohesin depleted mitotically arrested cells, CH, (bottom), for selected regions on (left) ChrXV (0-330 kb (CENXV is at 330 kb)) and (right) the post-rDNA region of ChrXII (660 kb to 940 kb). (d) Zoom into log2 ratio of CH over C contact maps for selected regions on (left) ChrXV (0-330 kb (CENXV is at 330 kb)) and (right) the post-rDNA region of ChrXII (660 kb to 940 kb). Arrows indicate a prominent track of cohesin-dependent contacts seen also in Supplementary Fig. 3a). All contact maps shown were assembled from two independent experiments.