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Fig. 3. Cytochalasin D disrupts the association of cortactin with dynamin 2 at endocytic sites. (A) NIH3T3 cells were treated with DMSO or 10 µM cytochalasin D in serum-free medium. After 1 hour of treatment, cells were treated with 20 µg/ml biotin transferrin and incubated for 10 minutes. In a parallel experiment, cytochalasin D was removed washed once with serum-free medium and the cells were incubated for additional 1 hour and followed by adding biotin-labeled transferrin. The treated cells were fixed, permeabilized and stained for dynamin 2 (green) and cortactin (red). The boxed regions shown in panels a, b and c were magnified and are presented in panels a', b' and c', respectively. N, nucleus. Arrows in a' indicate typical colocalization of dynamin and cortactin. Magnification x1000. (B) Colocalization of endogenous cortactin and dynamin 2 in NIH3T3 cells was quantified using Optimas 5.2 image analysis software. The data shown are the mean±s.e.m. (n=20).
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