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. 2014 May 1;25(9):1532–1542. doi: 10.1091/mbc.E13-10-0600

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© 2014 Miyamoto 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 4: — ACAP2 is inactivated following differentiation in FBD-102b cells. (A) The plasmid encoding myc-tagged ACAP2 was transfected into 293T cells and lysed to be used in an affinity precipitation to detect overexpressed, active ACAP2 using various forms of guanine nucleotide–binding recombinant Arf6. The GTP form of Arf6 only binds to ACAP2. Immunoblot for total ACAP2 is also shown. (B) Cells were allowed to differentiate for 0–3 d and lysed to be used in an affinity precipitation of active ACAP2 with recombinant GST-tagged GTPγS-bound Arf6. Immunoblots for total ACAP2 and β-actin are also shown. (C) Bands corresponding to each active ACAP2 protein were scanned and semiquantified (n = 4). Data were evaluated with one-way ANOVA (*, p < 0.01; **, p < 0.05).