Fig. 4. Hit validation.

a Selected hits were confirmed by dose–response DUB-ABPP over 100, 50, 10 µM. b Overall, we observed high confirmation rates and excellent agreement for assays spanning compound binding in mass spectrometry (red), western blot (green), and enzyme inhibition (blue) for library hits targeting UCHL1, UCHL3, USP7, USP28, USP48 and VCPIP1. An outlier (XL-9872-111B) for which we observed poor agreement across assay types is shown on the rightmost column. Data for each DUB is shown in a row (6 DUBs total) and data for each compound is shown in a column (24 compounds total). For individual compound structures and primary screen activity, please refer to Supplementary Data 3. c In a competition experiment against a desthiobiotinylated analog, F-70 shows dose–response binding to only 4 proteins among 1074 proteins detected with a 1% FDR cutoff as measured by a two tailed Z-test. Top DUB hits are highlighted in red. Source data are provided as a source data file.