Online supplemental material

Fig. S1 shows additional consequences of Ska complex depletion on mitotic fidelity, prevalence of merotelic KTs in Ska-depleted cells, comparable Ska1 knockdown efficiencies in HeLa S3 and HeLa K cells upon double and single siRNA treatment, respectively, and confirmation of the effects of Ska complex depletion on KT-MT dynamics with a siRNA against an alternative Ska complex subunit. Fig. S2 shows reduced Dsn1-pS100 and H3-pS28 levels in Ska-depleted cells, confirmation of the effect of Ska depletion on H3-pS10 with a panel of alternative siRNAs, and the FRET data from Fig. 2 (L and N) normalized to control and ZM cells. Fig. S3 shows that CPC protein levels and complex formation remain unaffected in Ska-depleted cells, that neither cohesion fatigue nor Haspin kinase inhibition impairs H3-S10 phosphorylation despite displacement of centromeric Aurora B, and that MCAK KT and H3-pS10 levels are not restored after ectopic centromere enrichment of Aurora B in the absence of Ska. Fig. S4 shows GFP-Ska1 fusion protein levels at KTs in the stable cell lines used in Fig. 4 and the dependence of Aurora B activity toward H3-S10 on spindle MTs. Fig. S5 shows purification of recombinant proteins used for in vitro kinase assays in Fig. 5 and additional control experiments related to Fig. 5. Table S1 shows the double exponential decay curve fitting parameters and sample sizes analyzed, related to Fig. 1 (F–I).