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Fig. 6. Colocalization of CAP1 with cofilin, and analysis of factors that determine its localization.

(A,B) Spreading mouse C3H-2K fibroblasts were fixed and incubated with a rat anti-CAP1 IgG and a rabbit antibody to actin (A) or cofilin (B), followed by labelling with a Cy3-labeled anti-rat IgG and a fluorescein-labeled anti-rabbit IgG. The lamellipodia are indicated by arrowheads, whereas the ruffling areas are indicated by arrows.

(C) HA-tagged CAP1-NT was transiently expressed in C3H-2K cells (panels a-d) by lipofection. HA-tagged CAP1-CT (panels e and f) or HA-tagged CAP2-NT (panels g and h) were expressed as well. Transfected cells were split onto coverslips, and the spreading cells were fixed and reacted with a mouse anti-HA and a rabbit antibody to actin (panels a, b, e and f) or to cofilin (panels c, d, g and h), after which they were stained with a Cy3-labeled anti-mouse IgG and a fluorescein-labeled anti rabbit IgG. The lamellipodia are indicated by arrowheads (for CAP-NTs) or arrows (for CAP1-CT). The white bar represents 10 µm.





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