Effects of anti-PLSCR1 on anchorage-independent colony formation, migration, and invasion of CRC cells. (A) The anchorage-independent colony-forming ability of HT29 cells treated with isotype-control IgG or NP1 was compared using the soft-agar cell transformation assay. Scale bar = 1.0 mm. (B) Migration of HT29 and HCT116 cells treated with NP1 is impaired compared to cells treated with isotype-control IgG. Cell-covered areas were quantified using the Wimasis image analysis platform. (C) Invasion assays with HT29 and HCT116 cells. The chamber was incubated at 37°C for 48 hours and cells that had invaded the polycarbonate membrane to the lower surface were quantified by counting five random microscope fields. Representative photomicrographs of filters (left panel) and quantitative analysis of the assay with HCT116 cells (right panel) are shown. Each bar represents the mean ± SD calculated from three independent experiments.