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. Author manuscript; available in PMC: 2014 Apr 18.
Published in final edited form as: Chem Biol. 2013 Apr 18;20(4):604–613. doi: 10.1016/j.chembiol.2013.04.001

Figure 3.

Figure 3

Potency and Selectivity of 2-D08. (A) IC50 for 2-D08 is measured to be 6.0 µM using the mobility shift assay. Reactions were performed at varying concentrations of 2-D08 and quenched with EDTA after 90 minutes. Conversion was measured using the Caliper EZ Reader II system. Values represent the mean of three replicates. (B) 2-D08 dose-dependently inhibits the sumoylation of IκBα with SUMO-1, SUMO-2 (C), and SUMO-3 (D) as observed by Western blot using anti-IκBα (C-21) antibody. Sumoylation of full-length protein IκBα (human, recombinant) was performed with recombinant SUMO-1, SUMO-2, or SUMO-3 in the presence/absence of 2-D08 at room temperature for 90 minutes and sumoylated IκBα level was measured by Western blot analysis with an anti-IκBα.