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Fig. 6. Clustering of SNAKA51-bound {alpha}5 integrin induces integrin translocation out of focal adhesions and across the cell surface. Fibroblasts were plated on glass coverslips and incubated overnight in fibronectin-depleted medium with cycloheximide. Cells were labeled with antibody for 20 minutes, using (a) SNAKA51, (b) SNAKA52 or (c) mAb11. Unbound antibody was rinsed off and the cells were either fixed (no chasing) or the cell-bound antibody was clustered with goat anti-mouse or anti-rat IgG and incubated for a further 30 minutes ('30' chasing) before fixation. For unclustered {alpha}5 integrin, cells were stained with anti-mouse or anti-rat IgG (green). For clustered {alpha}5 integrin, cells were stained with anti-goat IgG (green). In addition, all samples were stained with anti-{alpha}V antibody L230 (red) as a focal adhesion marker. Bar 20 µm.





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