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. 2011 May 23;6(5):e19720. doi: 10.1371/journal.pone.0019720

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Chew et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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In vitro ubiquitination reaction was carried out with synphilin-1 as a substrate in the absence of parkin (i.e. –E3) or in the presence of MBP-parkin (FL) or IBR-R2 in combination with either UbcH7, Ubc13 or no E2 (as indicated). Parkin-mediated ubiquitination of synphilin-1 was analyzed by means of anti-synphilin-1 immunoblotting (top panel) whereas parkin-mediated self ubiquitination was analyzed by means of anti-parkin immunoblotting (bottom panel).