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. 2016 Nov 28;45(4):1793–1804. doi: 10.1093/nar/gkw1162

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — AR K311 is not the sole target of MDM2-mediated ubiquitination. (A) HEK293T cells were transfected as indicated and AR with interacting partners were immunoprecipitated followed by immunoblotting. (B) HEK293T cells were transfected as indicated followed by denaturing immunoprecipitation and immunoblotting. (C) Western blotting of AR DBD/H/LBD with annotated monoubiquitination (AR-Ub1) and di-monoubiquitination (AR-diUb1) by MDM2 following in vitro ubiquitination assay. Reactions contained UBE1, UBE2D3, and UbK0 in the absence or presence of ATP to activate the ubiquitination reaction.