Fig 5. (A) Soft agar colony formation assays were performed on each clone as a measure of cell-adhesion growth. IH-selected clones demonstrated an increase in the number of colonies relative to passage control clones. Metastatic MDA-MB-231 cells were seeded as a positive control. (B) Passage control MCF01A clones (top) and IH-selected clones (bottom) were imaged using brightfield microscopy. While control clones exhibited smooth borders indicative of an epithelial phenotype, IH-selected cells migrated apart from adjacent cells, indicating an increase in migration as well as EMT. (C) As a measure of migratory capacity, passage control MCF10A clones (in grey) and IH-selected MCF10A (in black) were seeded within an xCELLIGENCE CIM plate, which measures in real time the number of cells passing through a barrier from a low-serum well to high-serum well. Measurements taken over the span of 24 hours indicates IH-selected clones have increased migratory rates (D) Proliferation rates of each of the four clones. Cell counts were performed daily. IH-selected clones exhibited reduced proliferation relative to passage control clones. (E) Oxygen consumption and proton production (a proxy for glycolysis) were measured for each clone with the Seahorse XF Analyzer. The ratio of oxygen consumption to proton production was increased in IH-selected clones.