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. 2015 Jul 27;112(32):9872–9877. doi: 10.1073/pnas.1505923112

Fig. S3.

Fig. S3.

The ubiquitin-loading status of UbcH5b affects its ability to interact with E6AP. UbcH5b–ubiquitin conjugates (20 µM) were incubated in the absence or presence of increasing concentrations of the HECT domain of E6AP as indicated for 5 min at room temperature. The mixtures were fractionated by size exclusion chromatography (SEC) and the fractions were analyzed by SDS/PAGE followed by Coomassie staining. Relative fraction numbers are indicated. Running positions of the HECT domain and the UbcH5b–ubiquitin conjugate are marked by an arrowhead and an asterisk, respectively.