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. Author manuscript; available in PMC: 2014 Oct 9.
Published in final edited form as: Clin Cancer Res. 2013 Nov 5;20(2):372–381. doi: 10.1158/1078-0432.CCR-13-1252

FIGURE 1.

FIGURE 1

SCCRO5 interacts with components of the neddylation pathway. A, Western blot analysis of the products of GST-SCCRO, GST-SCCRO5, and selected GST-SCCRO5 mutants after pull-down assays from HeLa lysates probed with the indicated antibodies, which shows that, like SCCRO, SCCRO5 binds to CAND1, Cul1, Cul2, Cul3, and ROC1. Mutations in the PONY domain (SCCRO5_D225N and SCCRO5_E226A), but not the N-terminal NLS (SCCRO5_Δ1-46), result in loss of binding. The dividing line between lane 1 and lane 2 indicates the position of omitted lanes from the same gel. B, Western blot analysis of Ubc12 after GST and GST-SCCRO5 pull-down assays on products from a thioester reaction (lane 1), which shows preferential binding to Ubc12~Nedd8, even in the presence of excess free Ubc12. C, Western blot analysis of Ubc12 after pull-down assays on products of thioester reaction, which shows that GST-SCCRO5 and PONY and NLS mutants bind equally to Ubc12~Nedd8.