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Study of E6–paxillin interaction by cell fractionation and cytochalasin D treatment. COS cells transfected with FLAG-tagged E6 were fractionated into cytosolic (fraction a), membrane (fraction b), and nuclear (fraction c) fractions. Half of the cells were treated with 2 μM cytochalasin D (Sigma) for 1 h before harvest. Fractionated lysates were immunoprecipitated using a paxillin mAb (Chemicon) or the M2 FLAG antibody and blotted for paxillin or FLAG–E6 as indicated.
