Fig. 5. Effect of olomoucine on Src activation and F-actin in scratch-wounded cultures. Confluent cultures of A6(1) cells were scratch wounded then cultured for 12 hours with or without 15 µM olomoucine. After 12 hours, cultures were double stained with antibody for active Src (pY416) and rhodamine-phalloidin. (A) Active Src is seen in a band of cells along the wound edge. (B) In the presence of olomoucine, the distribution of active Src is altered. Intense staining is seen immediately adjacent to the wound edge. (C) A higher magnification view of cells shown in (A). Active Src is localized to the cell periphery (arrow) and the perinuclear region (arrowhead). (D) Higher magnification of cells shown in (B). In the presence of olomoucine, immunostaining of active Src is enhanced in lamellipodia along the cell periphery (arrow) and diminished in the perinuclear region (arrowhead). (E) Rhodamine-phalloidin staining of same cells as (C) shows that F-actin is organized into thick bundles that appear as large points or elongated ovals in the optical section (arrows). (F) Rhodamine-phalloidin staining of cells shown in (D). In the presence of olomoucine, actin staining is more diffuse and sectioned actin fibers are narrower. Lamellipodia are not strongly stained for F-actin (arrows). Bar, 250 µm (A,B), 25 µm (C,D).