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Fig. 8. Autocrine stimulation leads to elevated cell migration speed. Cell migration was measured in a sparse-labeled monolayer high-throughput assay. Parental, ECT and TCT cells were starved for 15 hours and then switched to fresh serum-free media, 0.2 nM EGF, 2 nM EGF, 10 µg/ml mAb225 or batimastat (0.12, 0.33, 0.86 or 10.00 µM) 
1 hour before imaging. Fluorescence images were acquired at 15-minute intervals for 8 hours and imported in Imaris for single-cell tracking of the labeled cells. Cell paths were analyzed in Matlab. (A) Cell paths are shown starting at each origin. (B) Average cell migration speed under each condition. (C) The average cell speed was measured for all cell types when stimulated with exogenous EGF while ligand cleavage at the cell surface was inhibited with batimastat. After 14 hours of starvation in serum-free media, the cells were treated for 2 hours with 10 µM batimastat and then switched to fresh batimastat alone or with 2 nM EGF. Cell speed is compared to that measured under serum-free media alone for each cell type. Error bars represent s.e.m., and n >200 cells per condition.
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