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. 2011 Sep 1;22(17):3206–3217. doi: 10.1091/mbc.E11-02-0145

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© 2011 Aoki 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 3: — Phosphorylation of Ser-114 on Atg32 is critically important for the Atg11–Atg32 interaction. (A, B) Yeast two-hybrid analysis between Atg11 and the indicated Atg32 mutants. (C) The atg11Δ atg32Δ strains expressing PA-tagged Atg32^WT, PA-tagged Atg32^S114A, or PA-tagged Atg32^S114T and HA-tagged Atg11 under the control of the CUP1 promoter were grown in SMD medium until the mid–log phase and then starved in SD-N for 1 h. PA-Atg32 was precipitated using IgG–Sepharose from cell lysates. Top two, an immunoblot of total-cell lysates. Bottom two, the IgG precipitates, which were probed with anti-HA and anti-PA antibodies.