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Effect of C25-140 on various E3 ligases/E2 enzyme reactions. A, cIAP1 generating Lys63-ubiquitin chains was also inhibited by C25-140. B, MDM2 and TRIM63 generating Lys48-ubiquitin chains were not impaired by C25-140. C, RNF4, a SUMO-targeted ubiquitin ligase, was also not affected by C25-140. D, activity of the HECT E3 ligases ITCH and E6AP was not inhibited by C25-140. The substrates and the compound concentrations are indicated; assays were purchased from Boston Biochem.
