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To take sequential pictures and make movies, transfected cells were plated on 35-mm glass bottom culture dishes (MatTek, Ashland, MA) precoated with 2 μg/ml CS-1, scratch wounded, and placed on an open chamber with temperature controller (Harvard Apparatus, Holliston, MA). Plates were washed twice with PBS to remove unbound cells at designated time points after scratch wounding, and cells were viewed on a phase contrast microscope (Nikon, Melville, NY) and photographed with an attached camera (Diagnostic Instruments, Sterling Heights, MI). The cells were washed twice with phosphate-buffered saline (PBS) and then cultured in media containing 10% FBS in a tissue culture incubator. Briefly, transfected cells were plated onto 24-well tissue culture plates coated with 10 μg/ml human plasma FN (Roche Diagnostics) or GST-CCBD, or 2 μg/ml CS-1.
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Interestingly, it has recently been shown that regulation of integrin function contributes to the complex behaviors of neural crest cells (Strachan and Condic, 2003, 2004). For example, if a cell expresses both α4β1 and α5β1 integrins, which bind to different regions of FN, expression https://www.hierrosyacerosconil.com/ of ADAM12 could promote use of α5β1 for migration. Results presented in this study suggest that ADAMs may also influence cell migration in a manner independent of proteolytic activity. For example, ADAM proteases may shed factors that stimulate or direct cell migration (Maheshwari et al., 2001; Mechtersheimer et al., 2001). The patterns of inhibition are consistent with the integrin binding profiles of the respective ADAM disintegrin domains on cells with activated β1 integrins (Bax et al., 2004) (Figure 1; our unpublished data).
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