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. 2011 Feb 1;22(3):315–326. doi: 10.1091/mbc.E10-04-0360

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© 2011 Tan 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 7: — The Db1 and GxEN degrons direct Ama1p association. (A) An ama1Δ mutant (RSY562) harboring different Ama1p and Cdc20p expression plasmids (Ama1p^CBΔ/IRΔ, pKC3048; Cdc20p, pMSC8; Ama1p^CBΔ/IRΔ, pKC3048; Cdc20p^CBΔ/IRΔ, pKC5022; vector, pRS424) as indicated was induced to enter meiosis and cells were harvested for analysis at 9 h, when both AMA1 and CDC20 are expressed (Cooper et al., 2000). Immunoprecipitation and Western blot analyses were conducted to detect the presence of both proteins (top and middle) or coimmunoprecipitation (bottom). (B) Coimmunoprecipitation experiments performed as described in (A) were repeated except with the inclusion of the Cdc20p^DB1/GxEN-18myc (pKC5023) expression plasmid. The no-antibody mock immunoprecipitation is represented by [ ].