ILK knockout in combination with bosutinib treatment causes cell adhesion defects. A, Western blot analysis of cell lines grown in 2D. Data are representative of three independent experimental replicates. B, Adhesion challenge: IncuCyte cell confluence was normalized to DMSO PX459v2. DMSO, solid fill; bosutinib, clear fill. C, 2D 8-point dilution for half-inhibitory concentrations (EC50) for PND-1186. Endpoint quantification using normalized cell counts form Hoechst-stained images. D, Cell-by-cell mask and a “rounded cells” classifier whereby area <600 μm2 and eccentricity <0.65, one hour post drug addition. E, IncuCyte S3 cell-by-cell quantification for MDA-MB-231 cells plated at 8,000 cells per well. Arrow, drug addition. A bosutinib EC20 (0.9 μmol/L) concentration was used in all experiments. ***, P < 0.001.