Confirmation of positives from the high-content cell migration screen. A. Multiparameter concentration-response confirmation in the Oris™ Pro assay.

<p>MDA-MB-231 cells were treated for 48 h with ten point, two-fold concentration gradients of test agents. Cells were stained with Hoechst 33342 and analyzed for cell migration, cell density, and condensed nuclei as described in Materials and Methods. Data are the averages ± SE from quadruplicate determinations and are from a single experiment that has been repeated twice. B. and C. Single-cell motility assay. Agents that showed selective, concentration-dependent inhibition of cell migration in the primary assay format were tested in a single-cell motility assay. Two compounds significantly reduced cell migration velocity compared with vehicle control. Box, 25th and 75th percentiles, whiskers, 10th and 90^th percentiles; line, median. Data are the combined values from two independent experiments, each comprising 28 individual cells. Rose plots in C. illustrate motility patterns of individual cells. Each line represents the trajectory of a single cell over a period of 48 h.