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. 2017 Nov 17;292(52):21282–21290. doi: 10.1074/jbc.C117.815845

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 2. — A, FOS stability assay on HUVECs stably expressing FOS or FOSΔ. B, ubiquitin assay of cells transfected with the indicated constructs together with 10× HIS epitope-tagged ubiquitin. C, left panel, FOS stability assay on HUVECs stably expressing FOS in the presence or absence of MLN7243. Right panel, ubiquitin assay of cells transfected with the indicated constructs and cultured in the presence or absence of MG132 and MLN7243. IP, immunoprecipitation. D, in vitro translated FOS proteins were incubated with purified 20S proteasomes for the shown time course (minutes). 20S protein levels were determined by Western blotting using an antibody directed against PSMA1. 20S proteasome activity was independently quantified using the suc-Leu-Leu-Val-Tyr-AMC peptide (as shown in E). E, experiment performed as in D.