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. 2013 Jun 1;24(11):1676–1687. doi: 10.1091/mbc.E12-10-0772

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© 2013 Dui 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: — dSkp2 interacts with Dap and regulates Dap protein level in Drosophila tissues. (A) CoIP analysis detecting dSkp2-Dap interaction. Extracts were prepared from the adult heads of flies with the indicated genotypes: GMR>HA-dSkp2 (lanes 1, 4, 7, and 10), GMR>4xMyc-dap (lanes 2, 5, 8, and 11), and GMR>HA-dSkp2 + GMR>4xMyc-dap (lanes 3, 6, 9, and 12). The immunoprecipitates were pulled down with the anti-Myc antibody, and the anti-HA antibody was used in IB to detect the presence of HA-dSkp2. (B) dSkp2 regulates Dap stability. Extracts were prepared from the adult heads of flies as in A. Total protein level in the extracts was detected in IB using the indicated antibodies. Tubulin (lanes 7–9) is loading control. (C) dSkp2 knockdown in Drosophila eyes leads to accumulated Dap proteins in tissue extracts. Extracts were prepared from the adult heads of flies with the indicated genotypes: GMR>4xMyc-dap (lanes 1 and 3) and GMR>4xMyc-dap + GMR>dSkp2^GD5142 (lanes 2 and 4). Myc-tagged Dap protein level was determined by Western blotting using the Myc antibody. Tubulin (lanes 3 and 4) represents loading control.