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).为优化基于NGI-1的STT3A抑制剂，研究人员采用DDT细胞对约1500种NGI-1衍生物的STT3A抑制活性开展筛选。将细胞以3000个细胞/孔（40 μL）的密度接种于384孔板中。经过20小时的贴壁培养后，向细胞中加入100 ng/mL的Wnt3a蛋白（Wnt3a protein，5 μL）与8 μM的NGI-1衍生物（5 μL）进行共处理。共处理24小时后，通过添加10 nM的AP1903诱导细胞凋亡。AP1903诱导16小时后，使用20 μL的8%多聚甲醛（PFA）固定细胞，并通过EnSight成像系统（珀金埃尔默，PerkinElmer）进行成像。