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. 2013 Aug 1;24(15):2378–2388. doi: 10.1091/mbc.E12-12-0860

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© 2013 Zlatic 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 clathrin-binding box of AP-3 β1 is dispensable for AP-3–clathrin coisolation. Pearl (Ap3b1^pe/pe) fibroblasts or Pearl fibroblasts rescued with full-length Ap3b1 (pβ3A), Ap3b1 containing a triple-alanine mutation in the clathrin box (pβ3A₈₇₁AAA), a deletion of the clathrin box in Ap3b1 (pβ3AΔ _807-831), or a truncation of Ap3b1 eliminating the entire ear domain containing the clathrin box (pβ3A₈₀₇Stop) were treated with the cross-linker DSP. Lysates were subjected to coimmunomagnetic isolations directed against AP-3 δ (lanes 1–10). All rescue lysates from DSP-treated cells show increased coisolation of clathrin heavy chain (immunoblot [IB]; clathrin lanes 4, 6, 8, and 10) compared with unrescued lysate treated with DSP (IB, clathrin lane 2). Input represents 5% of immunomagnetic isolation load (lanes 11–15).