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Fig. 9. Confluent cultures of A6(1) cells were scratch wounded then cultured with or without the Src inhibitor PP1 or the inactive analog PP3. After 24 hours, cultures were double stained with antibody for phosphorylated Cdk5(pY15) (A-C) and rhodamine phalloidin (D-F). (A) In the presence of PP3, Cdk5(pY15) immunofluorescence was observed in a band of cells along the wound edge. (B) In the presence of PP1, little or no Cdk5(pY15) immunofluorescence was detected along the wound edge. (C) Omission of the primary antibody to Cdk5(pY15) showed no detectable immunofluorescence. (D) Rhodamine-phalloidin staining of cells shown in (A) showed polymerization of F-actin along the wound edge, with diffuse cytoplasmic staining and formation of numerous lamellipodia (arrows). (E) Rhodamine-phalloidin staining of cells shown in (B) showed that the actin cytoskeleton was primarily cortical (open arrow) and few lamellipodia were formed. (F) Control cultures incubated without PP3 or PP1 were indistinguishable from those incubated with PP3. Numerous lamellipodia were observed (arrows).





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