Spheroid and monolayer plating schema and screen design.

<p><b>A</b>, one spheroid per well was formed in 96-well round bottom ultra-low attachment plates. Brightfield micrographs of typical BT-474 and MCF-10A spheroids are shown 48 hours after plating, <i>bar,</i> 200 microns. <b>B</b>, the 2D monolayer cultures were set using the same cell number as in the round bottom plates (3000 cells/well for BT-474 or 1000 cells/well for MCF-10A) in flat bottom, tissue culture-coated 96-well plates. Fluorescence images are shown of typical BT-474 and MCF-10A monolayers 48 hours after plating, stained for actin (red) and DNA (blue), <i>bar</i>, 200 microns. <b>C</b>, schematic of screen design.