Screen and Characterization data for NGI-1 derivatives related to the study "STT3A is essential for Wnt signaling and represents a target for cancers driven by RNF43 deficiency"

To optimize NGI-1–based STT3A inhibitors, approximately 1,500 NGI-1 derivatives were screened for their STT3A inhibitory activity using DDT cells. Cells were seeded in 384-well plates at a density of 3,000 cells/well (40 μL). After a 20-hour adhesion period, cells were co-treated with 100 ng/mL Wnt3a protein (5 μL) and 8 μM NGI-1 derivatives (5 μL). Following 24 hours of co-treatment, apoptosis was induced by adding 10 nM AP1903. Sixteen hours after AP1903 induction, cells were fixed with 20 μL of 8% PFA and imaged using the EnSight system (PerkinElmer).