Journal: PLoS ONE

Article Title: Endothelial Induced EMT in Breast Epithelial Cells with Stem Cell Properties

doi: 10.1371/journal.pone.0023833

Figure Lengend Snippet: A , D492M show increased migration compared to D492. Increased migration was seen at the two cell concentrations (1×10 4 and 2.5×10 4 ). B , D492M grow anchorage independently. In 0.5% soft agar D492 cells fail to form colonies. In contrast, the D492M cells are able to grow, indicating acquisition of anchorage independent growth. In monolayer culture (right) D492 cells are contact inhibited while D492M piles up in the culture flask indicating lack of contact inhibition (arrows). Counterstain hematoxylin. Bar 100 µm. C , D492M cells are CD44 high ,CD24 low consisting with the breast cancer stem cell phenotype. D492 contain a subpopulation (19%) of cells that are CD44 high , CD24 low . This population increases to 70% in the D492M cell line. D , D492 and D492M differ in their ability to form mammospheres. Both D492 and D492M can generate colonies in mammosphere assay, however, D492M generates more and larger (>100 µm: 1.7 fold; >150 µm:1.5 fold) mammospheres than D492 cells. E , D492M cells show delayed chemically induced apoptosis. D492 and D492M show distinct responses to Camptothecin, an apoptosis inducing agent. D492 cells underwent immediate apoptosis and showed cell survival under 40% on day 2 while having no effect on D492M. On day 3 D492M cells showed cell survival of 60% where only few D492 cells were left. Data shown as AVG number of cells per field (A,E) or AVG number of colonies (B,D) +SEM in three experiments. *p<0.05; **p<0.01.

Article Snippet: Cells were incubated for 20 minutes with fluorochrome-conjugated antibodies against CD44 (clone IM7, BD), CD24 (clone ML5, BD) or isotype-matched controls, subsequently washed and resuspended in PBS with 4% formaldehyde (cell-fix).

Techniques: Migration, Inhibition