SHCBP1, PLK1, and MISP gene were cloned into the pETDuetM-6HIS-GST-pp vector. GST-tagged SHCBP1, His-tagged PLK1, and His-tagged MISP proteins were expressed in the Escherichia coli strain BL21 (DE3) strain and GST pulldown assay was conducted, using a pierce GST protein interaction pulldown kit (Thermo Scientific). Input and eluate samples were resolved using SDS–PAGE and analyzed by Coomassie blue staining or immunoblotting.
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