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. 2022 Jul 1;221(8):e202108156. doi: 10.1083/jcb.202108156

Figure 1.

Figure 1.

CPC–Sgo1 forms a robust complex in vitro. (A) Schematic diagram depicting the domain architecture of Sgo1 and the Borealin, Survivin, and INCENP subunits of the CPC. CPCISB (INCENP1–58, Survivin full length, and Borealin full length) is highlighted in the red box. (B) SEC profiles and corresponding Coomassie-stained SDS-PAGEs for the analysis of Sgo11–415 (blue) and CPCISB10–280 (INCENP1–58, Survivin, and Borealin10–280; dark gray) complex formation (red). 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. Elution volumes of peak fractions are indicated above the chromatogram peaks. (C) Isotherms for Sgo11–415 interaction with CPCISB10–280 (40 μl of 50 μM CPCISB10–280 was injected into 200 μl of 5 μM Sgo11–415). The ITC experiment was performed with 16 × 2.5-μl injections (0.5 μl first injection), 180 s apart, at 20°C. Top panel shows the raw ITC data; bottom panel shows the integrated heat data corrected for heat of dilution and fits 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 F1.