Nischarin decreases the attachment of breast cancer cells. A, The 231 (n = 6) and 231 Nisch (n = 6) cells were seeded onto a standard tissue culture surface (2D), fibronectin, or collagen for 48 hours. Cell proliferation was determined by the MTT assay. B, Nisch+/+, Nisch+/−, and Nischr−/− cells were seeded onto a tissue culture surface (2D), fibronectin, or collagen for 48 hours. Cell proliferation was determined by the MTT assay. C, The 231 (n = 18) and 231 Nisch (n = 21) cells were seeded on 10 μg/mL fibronectin for 24 hours and stained with phalloidin. The average area and aspect ratio were calculated using ImageJ (inset with a zoom is shown). Aspect ratio = major axis/minor axis. D, Vinculin immunofluorescence staining of 231 and 231 Nisch cells (inset with a zoom is shown). Number of vinculin-positive focal adhesions (FA) per area and percentage of area covered by focal adhesion was calculated using CellProfiler. *, P < 0.05; **, P < 0.01; ***, P < 0.001.