Figure S2.

Sgo1–Survivin interaction is essential for CPC–Sgo1 complex formation. (A–C) SEC profiles (top) and corresponding representative SDS-PAGEs stained with Coomassie (bottom) for the analysis of Survivin and Sgo1_1–415 interaction (A), Survivin_1–116 and Sgo1_1–415 interaction (B), and CPC_ISB10–280 containing different Survivin mutants and Sgo1_1–415 interaction (C). For easy direct comparison, control SDS-PAGE and chromatogram corresponding to Sgo1_1–415 are shown in Figs. S2 A and S4 C. A Superdex S200 10/300 GL (Cytiva) column pre-equilibrated with 25 mM Hepes, pH 8, 250 mM NaCl, 5% glycerol, and 2 mM DTT was used. (D and E) Isotherms for the analyses of CPC_{ISB 10–280} interaction with Sgo1_1–415 (D) and CPC_{ISB 10–280} interaction with Sgo1_{1–415 Nmut} (E). 40 μl of 50 μM CPC_{ISB 10–280} was injected into 200 μl of 5 μM Sgo1_1–415 or Sgo1_{1–415 Nmut}. The ITC experiments were performed with 16 × 2.5-μl injections (0.5 μl first injection), 180 s apart, at 20°C. Top panels show raw ITC data; bottom panels show integrated heat data corrected for heat of dilution and fitted to a standard 1:1 binding model (Malvern Instruments MicroCal Origin software, v1.3). DP, differential power; mAU, milli absorbance units. Source data are available for this figure: SourceData FS2.