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A previous study demonstrated that GFP-tagged α4 integrin behaves like untagged α4 integrin (Pinco et al., 2002). Transwell migration assays were performed as described previously (Liu et al., 1999), with the following modifications. After incubation for 2 h in a CO2 incubator at 37°C, confluent cell monolayer was scraped with a yellow Pipetman tip to generate scratch wounds. Scratch wound migration assays were performed as described previously (Pinco et al., 2002). As described previously (Pinco et al., 2002), green fluorescent protein (GFP)-tagged α4 integrin is functionally normal. GST-tagged proteins containing the disintegrin domains of ADAM12 (wild type and mutant), ADAM19 and ADAM33 were produced exactly as described previously (Tomczuk et al., 2003).
In this report, we studied the https://ethereumcode.io/ effects of four ADAMs on cell migration mediated by either the α4β1or α5β1 integrins. We show, for the first time, that ADAMs can selectively regulate integrin mediated cell migration. As seen in Figure 8, although ADAM17 had no effect on α4β1-mediated cell migration (Figure 8A), it significantly inhibited α5β1 integrin-mediated cell migration (Figure 8B). We next assessed whether other ADAM family members can selectively inhibit integrin-mediated cell migration. Our results so far indicate that ADAM12 inhibits α4β1 integrin-mediated cell migration in transwell and scratch wound assays. The D488A mutant therefore likely has a reduced effect on α4β1-mediated cell migration (Figure 5B) because its disintegrin domain has a reduced affinity for α4β1 integrin (Figure 5A).
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