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. 2014 Nov 1;25(21):3389–3400. doi: 10.1091/mbc.E14-06-1087

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© 2014 Ling 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 6: — Osh4 positively regulates the Sec2–Sec15 interaction. (A) The Sec2–Sec15 interaction is inhibited in osh4Δ cells and sac1Δ cells. Sec2-3xGFP was immunoprecipitated with anti-GFP antibody from wild-type, osh4Δ, or sac1Δ cell lysates. As a negative control, wild-type cells expressing nontagged Sec2 were included in the coimmunoprecipitation experiments. We loaded 0.5% of whole-cell lysates as input and processed the rest of the samples for coimmunoprecipitation. Sec2-3xGFP in the immunoprecipitates was detected with anti-Sec2 antibody. Coprecipitated Sec15-13xmyc was detected with anti-myc antibody. (B) Quantification of the Sec2–Sec15 interaction in wild-type, osh4Δ, and sac1Δ cells. Three independent experiments were performed. The intensity of the bands was quantified using ImageJ. The percentage of Sec15 in the immunoprecipitate was calculated and is indicated. Mean and SD of three experiments.