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His-tagged proteins were purified using Ni-NTA Agarose (Cube Biotech) and dialysed in phosphate-buffered saline (PBS) (10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 , 137 mM NaCl, 2.7 mM KCl, pH 8.5), apart from SemDAPH for which a pH of 6.0 was used

Vitrification and data collection For all importin complexes studied in this paper, typically 2.0 l of importin pre-bound to IBB, xIBB, IBB:Nsp1, Ran-GTP, Ran-GTP:RanBP1, or Ran-GDP:RanBP1 at 2 mg/ml final concentration were applied to a 300-mesh copper Quantifoil R 1.2/1.3 holey carbon grid (EMS) previously glow-discharged negatively for 60 s at 15 mA using an easiGlow (PELCO)

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341,482 Up to now, MSNs have made great research progress in many aspects of tissue engineering, especially in bone tissue engineering