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Beads were washed once with lysis buffer, once with lysis buffer supplemented with 500 mM NaCl, once again with lysis buffer, and incubated with clarified bacterial lysates containing recombinant WIPI3-His6 , moesin(1300)-His6, COPB2(1304)-MBP-His6, SNX27-His6 , or His6-VPS26 for 2 h at 4 C on a rotator