Broad Library Small Molecule Screening Results

Table 1: Primary screening of Broad Library compounds in DM480 raw dataTable 2: Secondary of a select Broad Library compounds from primary screeningMethods:Cells were plated in culturing media at a density of 8000 cells per well in 384-well plates (Griener Bio-ONE Glass-bottom SensoPlate, Cat No. 781896) and allowed to recover for 24hours before being treated with compounds libraries. The compound library consisted of a subset of 371 commercially sourced small molecules from the Broad Informer Set49, which are stored as 10mM DMSO stocks on Echo Certified Low Dead Volume plates (Labcyte) and transferred to assay wells using Labcyte Echo 550 acoustic dispenser (Labcyte, San Jose, CA). Following 24hr compound treatment, cells were fixed, and FISH was performed as described above. Assay quality was monitored by evaluating the Z’ metric of the proportion of foci containing cells calculated from the on-plate negative (DMSO) and positive (Chromocycin [1 µM]) controls, which showed a mean value of 0.53. Indicated a sufficiently robust screening assay.