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Fig. 7. Binding to Arp2/3 complex is required for cortactin to recruit to dynamin 2. (A) MDA-MB-231 cells expressing cortactin-GFP or Cort(1-23)-GFP were grown in 10% serum medium and immunostained with GFP (a and d) or dynamin 2 antibody (b and e) as indicated. The stained cells were examined by confocal microscopy. Magnified images corresponding to the indicated boxes in merged pictures (c and f) are shown in c' and f'. Two dynamin structures that were closely associated with cortactin-GFP in panel c' are indicated by arrows. Magnification x1000. (B) Quantification of colocalization of cortactin-GFP and endogenous dynamin 2 using Optimas 5.2 image-analysis software. The data shown represent the mean±s.d. (n=20). (C) Cells expressing cortactin GFP variants were incubated with bio-transferrin for 10 minutes, lysed and subjected to immunoprecipitation with GFP antibody and further analyzed for the presence of cortactin and dynamin 2 by immunoblot assay. A representative blot is shown in the inset. The positions for dynamin 2 and cortactin GFP proteins are indicated. The density of the bands on the blot was digitalized and quantified. The data shown are the mean±s.d. of two independent experiments.

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