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. 2014 Apr 28;289(23):16430–16441. doi: 10.1074/jbc.M113.541490

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Inhibition of HERC2-FBXL5 interaction stabilizes FBXL5. A, HEK293T cells transfected (or not) with various amounts (1×, 2×, 4×) of an expression vector for FLAG-HERC2(2930–3327) were subjected to immunoblot analysis with antibodies to the indicated proteins. B, HEK293T cells transfected with expression vectors for FLAG-FBXL5(WT), FLAG-FBXL5(PE/AA), or FLAG-FBXL5(Δ257–297) were exposed to cycloheximide (100 μg/ml) for the indicated times and then subjected to immunoblot analysis with antibodies to FLAG and HSP90. The percentage of the FLAG-tagged proteins remaining after the various incubation times was quantitated by image analysis.