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The second basic cluster of Bag6 is necessary and sufficient for binding karyopherin α 2. (A) The putative bipartite nuclear localization sequence of Bag6. The serine and leucine mutations introduced in this study are highlighted. (B) Recombinantly purified Bag6C131–6xHis⋅Ubl4A (500 pmol) was incubated with twofold excess GST⋅TRC35 or MBP⋅KPNA2 for 20 min at room temperature. Ni-NTA beads were used to capture purified Bag6C131–6xHis⋅Ubl4A and bound factors.
