Fig. 2.

Notum activity-based occupancy assay was performed with FP-biotin (FP) (2 μM) and test compounds 1, 3, 5l and 24 (3 μM) for 30 min in conditioned media from HEK293S cells stably transfected with a Notum lentiviral construct. Relative occupancy was calculated by optical density of the fluorescent band, generated by streptavidin linked fluorophore to detect the level of biotinylation of Notum using Image Studio Lite 5.2, compared to the control-treated sample which was set to 1. N = 2 with S.D. UT, untreated.