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During the process of establishing contact inhibition of cell growth, cells form filopodia to establish cellular contact and to search for a suitable substrate. They are involved in the organization of cellular junctions and contain adhesion molecules such as integrins. Since some molecules involved in cell-substrate and cell-cell adhesion may be expressed in normal cells in a random distribution, their organization into specialized structures is considered as a normal function. On the other hand, malignant cells often lack structures related to contact inhibition. In order to observe the organization of cellular adhesion molecules in undisturbed conditions, we cultured NIH3T3 fibroblasts cells in a liquid surface culture system to induce contact inhibition. The filopodia formation during contact inhibition was observed by immunocytochemistry using anti-desmoplakin antibody (anti-DP) and by transmission electron microscopy. Anti-DP decorated all filopodia, indicating that they contain desmoplakin, a core component of the desmosomal plaque. The vinculin stained also the filopodia, indicating that they contain integrins. Upon immunoelectron microscopy, the anti-DP stained structures were found associated with the basal lamina with a zona occludens-like structure or at the level of the hemidesmosomal regions with a plaque-like structure. These results support the involvement of desmoplakin and integrins in normal contact inhibition.
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