Fig. 5. Inhibition of CDC34A-mediated in vitro ubiquitination by most potent isonipecotamide 2ab analogs.

(A) Gel-based assay for poly-ubiquitin chain formation. Concentrations tested ranged from 0.00084 to 50 μM in threefold increments. Experiment was performed twice, and representative gels from one experiment are shown. (B) Quantitation of poly-ubiquitin chain formation. Percent inhibition of poly-ubiquitin chain formation for each inhibitor concentration represents the mean from two experiments. (C) Comparison of TR-FRET EC₅₀ and ubiquitination IC₅₀ values. EC₅₀ values represent the mean ± SD (n = 3). IC₅₀ values represent the mean ± variance (n = 2).