Fig. 7. Translocation of Rok from cytosol to the membrane in neutrophils treated with colchicine or chemotactic peptide. (A,B) Neutrophils were preincubated for 15 minutes at 37°C without or with 10 µM taxol. The incubation was continued for another 5 minutes in medium (control) or with 10 µM colchicine (colch.) or with 10 µM lumicolchicine (lumicolch.). For the incubation with chemotactic peptides, cells were preincubated for 5 minutes at 37°C, followed by addition of 1 nM fNLPNTL (CP) and a further incubation for 15 minutes. Membrane and cytosolic fractions were subsequently prepared and analyzed by immunoblotting for the presence of Rok (arrow). (A) Immunoblot of a typical experiment; (B) Quantitative evaluation of A (mean±s.e.m. of three experiments; cells with colchicine: n=7). (C,D) Neutrophils were preincubated for 15 minutes at 37°C, without or with 10 µM taxol. The incubation was continued for another 15 minutes either in medium (control) or with 1 nM fNLPNTL (CP) and a further incubation for 15 minutes. The amount of Rok present in the membrane fraction was then analyzed by immunoblotting. (C) Immunoblot of a typical experiment (duplicates of each assay are shown); (D) quantitative evaluation of C (mean±s.e.m. of six experiments).

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