Fig. 1
From: Screening of breast cancer stem cell inhibitors using a protein kinase inhibitor library
Formation of mammospheres from MCF-7 cells and automated counting of mammospheres using NICE software. a Spheres derived from a single cell suspension of MCF-7 cells are visible in 7 days of culture with complete MammoCult™ medium. CD44 expression in mammospheres (gray) assessed by FACS analysis using FITC-anti-CD44 is higher compared to adherent MCF-7 cells (black). b The representative image of tumor spheres are shown with diameter of each measured by NIS-Elements BR4.4 software (Nikon, Tokyo, Japan). c Among mammospheres in the initial scanned image (Original), spheres larger than 50 μm in diameter are selected, counted and marked with a red crosshair (Counted) after histogram pitting using NICE software. The number of mammospheres counted by NICE software is comparable with the result of manual counting by naked eye under an inverted microscope. d Metformin (5 and 10 mM) significantly ameliorates the formation of mammospheres, which is demonstrated as a decrease in mammosphere formation efficiency (MFE) quantified by the NICE program and manual counting. The data shown are the mean ± SD of 3 independent experiments. *p < 0.05 vs. 0 mM. #p < 0.05 vs. 0 and 5 mM. The scale bar is 100 μm