Extended Data Fig. 2. Validation of primary HTS hits.

Candidate LYPLAL1 inhibitors and activators were analyzed by gel-based ABPP. Hit picks (10 μM) were incubated with 100 nM of purified mLYPLAL1 for 1.5 h at 37°C prior to the addition of FP-rhodamine probe (1 μM). After 1 h at 37°C, reactions were quenched, separated by SDS-PAGE and in-gel fluorescence scanned. LYPLAL1 activity was calculated relative to DMSO (100%). Designations above lanes correspond to compound location on the plate. Red asterisk denotes 4. Representative results from two independent experiments; similar results were obtained in both experiments. Uncropped gels are shown in Supplementary Fig. 6