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. 2020 Mar 5;39(7):e103234. doi: 10.15252/embj.2019103234

Figure 9. CAL1 can associate with CENP‐A/H4 heterodimer and CENP‐C simultaneously.

Figure 9

  1. (left panel) Normalised sedimentation coefficient distribution (c(s)) for CAL1841–979 (CAL1, blue), CENP‐C1264–1411 (CENP‐C, red) and their equimolar mix (complex, purple) all at 10 mg/ml, demonstrating a significant increase in s20,w0 consistent with the formation of a 2:1 complex. (right panel) Typical sedimentation equilibrium data for CAL1841–979 (CAL1, blue), CENP‐C1264–1411 (CENP‐C, red) and their equimolar mix (complex, purple) all at 10 mg/ml, demonstrating a significant increase in mass, consistent with the formation of a 2:1 complex. The data were fit with a single species model yielding masses of 20,253, 29,216 and 49,539 g/mol. The values reported in Fig EV5C are based on data acquired for a range of concentrations.
  2. SEC profile of His‐CENP‐C1264–1411 (red), His‐CAL11–160‐LL‐841–979/CENP‐A101–225/H4 (black) and His‐ His‐CAL11–160‐LL‐841–979/CENP‐A101–225/H4 mixed with molar excess of His‐CENP‐C1264–1411 (blue) and corresponding SDS–PAGE analysis of the fractions. Samples were analysed using Superdex 200 increase 10/300 in 20 mM Tris–HCl pH 8.0, 100 mM NaCl and 2 mM DTT.
  3. Schematic model of CAL1‐mediated loading of CENP‐A/H4 and CENP‐C binding at centromeres.