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. 2011 May 1;22(9):1575–1584. doi: 10.1091/mbc.E10-08-0716

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© 2011 Liu 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: — Interaction of Skp1 with Roy1 is required for its binding to Ypt52. (A) Schematic representation of Roy1 deletion mutants generated in the present study. (B) Yeast cells were cultured in YPD medium, and then the expression of Roy1 was induced in YPG medium for 12 h. Lysates were subjected to IP with anti-FLAG antibodies, and the resulting precipitates and total cell lysates were subjected to IB with the indicated antibodies. (C) The indicated cells were cultured in YPD medium to an OD₆₀₀ = 0.5 at 25°C and then at 37°C for 2 h. Lysates were subjected to IP and IB as described in B. (D) At 48 h after transfection with the expression vectors for the indicated proteins, 293T cells were harvested. Lysates were subjected to IP with the indicated antibodies, and the resulting precipitates and total cell lysates were subjected to IB with the indicated antibodies.