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. 2009 Dec;11(12):1318–1328. doi: 10.1593/neo.91084

Figure 5.

Figure 5

Self-renewal and mammosphere formation of Twist-expressing MCF-7 cells. MCF-7/Twist cells were stained with CD44 and CD24 antibodies and flow sorted to purify the CD44+/CD24-/low subpopulation. The purified MCF-7/Twist CD44+/CD24-/low subpopulation was propagated in culture, and the percentage of CD44 and CD24 cells was estimated by flow cytometry. (A) Histograms of chronological changes in the CD44+/CD24-/low subpopulation during a period of 13 generations. (B) qRT-PCR analysis of Twist, CD44, and CD24 transcript levels in the purified CD44+/CD24-/low versus CD44+/CD24+ subpopulations of cells. (C and D) Mammosphere formation by stable Twist-overexpressing cell lines, MCF-7/Twist, and MCF-10A/Twist. Top rows show phase-contrast photomicrographs of mammospheres at a magnification of x20. The lower rows show viability of the cells as indicated by the fluorescence of Calcein-AM stain. Data are represented as histograms on the right. Experiments were performed in quadruplicates and an average of five fields per well were counted.