Cell lysate (500 μg) was mixed with 5 μg of control IgG or CD13 mAb 452 overnight at 4°C with constant rotation, and proteins were immunoprecipitated with protein G–conjugated agarose beads (Invitrogen) by constant rotation for 2 hours at 4°C. The beads were washed four times in 1% NP-40 buffer containing protease and phosphatase inhibitors, and the precipitated protein was analyzed by immunoblot using anti-IQGAP1, anti-ARF6, or anti-EFA6 antibodies.

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