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. 2015 Jan 15;26(2):256–269. doi: 10.1091/mbc.E14-10-1451

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© 2015 Ren 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: — Identification of Cdc15_SH3 binding partners. (A) Summary of mass spectrometry results. Percentage sequence coverage and total spectral counts (TSCs) are shown. TSC is a combination of spectral counts from experiments using lysate from mitotic (Mito) and asynchronously (Asy) growing cells. The ability of GST-Cdc15_SH3 to pull out the indicated proteins is listed in the Pulldown column. (B) Recombinant GST or GST-Cdc15_SH3 on glutathione beads was added to protein lysates from rgf3-Myc₁₃ or spa2-FLAG strains and washed, and bound proteins were detected by immunoblotting with appropriate antibodies. Anti-Myc and anti-FLAG immunoprecipitates from the relevant strains served as positive controls. *: nonspecific bands and/or degradation products. (C) Heatmap of proteins identified in Rgf3-TAP and Spa2-TAP in prometaphase (Block, B) and after a 30-min release into anaphase (Block and Release, BR) by mass spectrometry. Only proteins that localize to the cell division site and were identified with >50 TSCs are listed. See Supplemental Table S2 for a complete list of all proteins identified. The values shown for each protein represent the relative abundance (normalized to bait) for each purification.