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. 2015 Feb 15;26(4):622–635. doi: 10.1091/mbc.E14-02-0740

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© 2015 Cleghorn et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 2: — Arrestins regulate cell migration and adhesion. (A, B) Adhesion was measured by plating cells on serial dilutions of FN (0.01–1.25 μg/ml) for 15 (A) or 30 (B) min. The data were analyzed by one-way analysis of variance (ANOVA) with arrestin type as the main factor, which was highly significant at 30 min. DKO cells showed a dramatic increase in their ability to adhere compared with WT cells, ***p < 0.001, **p < 0.01. Means ± SD from three experiments. (C) Adhesion of DKO cells expressing arrestin-2 + GFP, arrestin-3 + GFP, or GFP alone (controls). Cells were plated on 0.32 μg/ml FN. Means ± SD from 24 data points in three experiments. ***p < 0.001 compared with DKO. (D) Cells were plated in Transwell chambers coated with 0.32 μg/ml FN and allowed to migrate for 4 h. Cells were counted in six fields/chamber in each of four independent experiments. The data were analyzed by one-way ANOVA with cell type as the main factor, ***p <0.001. Insets, representative membranes postmigration. (E) Migration of DKO cells expressing arrestin-2 and GFP or arrestin-3 and GFP, or cells expressing GFP only (DKO and WT). Means ± SD from 5 fields/chamber from three independent experiments performed in duplicate analyzed by one-way ANOVA with cell type as the main factor. ***p < 0.001 compared with WT. DKO-Arr2, ^##p < 0.01, and DKO-Arr3, ^#p < 0.05, compared with DKO. (F) Arrestin expression in DKO cells was determined using arrestin-2– or arrestin-3–specific antibodies, with corresponding purified bovine arrestins (0.1 ng/lane) run as standards.