Fig. 3
Secondary CRISPR/Cas9 knockout screens using a focused library to target 599 candidate T-DM1 sensitivity/resistance genes. A Schematic of the secondary screens. B Cell growth for the control and T-DM1-treated cultures during the screens. Each condition was performed with duplicate cultures derived from a single (MDA-MB-361) or distinct (MDA-MB-453) founder libraries and the mean growth curves are plotted. C Emergence of resistance to T-DM1 at the end of the screens (day 32) in cultures that were treated with T-DM1 during the screens relative to untreated (naïve) cultures in MDA-MB-361 cells, as assessed by clonogenic survival of cultures exposed continuously to 1 or 1.4 nM T-DM1 over 21 days. D Volcano plot of the statistical significance of gene-level enrichment or depletion as a function of the median log2 fold-change in normalised read counts (T-DM1/control) of all sgRNA against each target (4 sgRNA per gene) in all replicate screens performed in MDA-MB-361 and MDA-MB-453 cells