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. Author manuscript; available in PMC: 2016 Oct 13.
Published in final edited form as: Nature. 2016 Apr 13;532(7599):398–401. doi: 10.1038/nature17433

Extended Data Figure 5. Free chains in trans do not stimulate USP14 activity for singleton conjugates.

Extended Data Figure 5

In vitro deubiquitination assays with single chain conjugates in the presence of free chains. ADP-Ptsm (4 nM) was incubated with Ubn-1KNCB1 (~110 nM) in the presence or absence of USP14 (80 nM), and with K48Ub4 (a), K48Ub2 (b), or free ubiquitin (c) at the indicated concentrations. Samples were analysed by SDS-PAGE and immunoblotted with an antibody to the HA epitope.