Expression of the “active” GIT1-Y246E/Y293E mutant increases the efficiency of migration.

<p>(<b>A</b>) COS7 cells transfected with GFP or with GFP-tagged GIT1 constructs were used for haptotactic cell migration towards fibronectin. Images show fields with cells migrated to the lower side of the filters coated with fibronectin. (<b>B,C</b>) Quantification of the number of cells migrated to the lower side of the wells. Bars are normalized mean values ±SEM from 12–19 fields from 2–3 independent experiments. **P<0.02. (<b>D–F</b>) Wound-healing assay. COS7 cells were transfected as in (<b>A</b>). Confluent monolayers were wounded and followed by time lapse imaging. (<b>D</b>) Fluorescent images (left column) showing GFP-positive transfected COS7 cells at time 0. The other columns show the phase contrast images of the same fields, at the beginning (t = 0) and the end (t = 6 h) of the assay. Bar, 100 μm. (<b>E</b>) Tracks of GFP-positive cells transfected with either GFP-GIT1 or GFP-GIT1-Y246E/Y293E. Tracks were taken during the wound healing assays and refer to an interval of 8 h. (<b>F</b>) Quantification of the euclidean distance covered by cells during the wound healing assay. Bars are mean values ±SEM from 90 cells from 3 different experiments. *P<0.05; ***P<0,002.