Fig. 3.

Nocodazole-induced increase in focal adhesion size in myosin-II-inhibited cells, but not in untreated cells. (A–D). Focal adhesions were stained with immunofluorescence against paxillin. The total area occupied by focal adhesions in nocodazole-treated cells (B) showed no significant difference from that in control cells (A). Cells treated with blebbistatin showed only dot-shaped focal complexes (C); large focal adhesions appeared upon nocodazole treatment (D). (E) Quantification of the average focal adhesion size was performed by manual segmentation. Similar results were obtained by staining for FAK and phosphotyrosine (pY). The increase in focal adhesions after nocodazole treatment in blebbistatin-treated cells was not observed in cells pretreated with a FAK-specific inhibitor PF 573228. Error bars indicate s.e.m. Scale bar: 25 μm. A total of at least 65 individual focal adhesions for each condition from at least four separate cells were analyzed for each condition. The timeline indicates the temporal sequence of drug manipulation.