Figure 5

TERT-siSFRP1 cells are anoikis resistant and exhibit CD44^high/CD24^low cell surface marker expression pattern. A, Cell culture dishes were coated with 1% agarose/DMEM, which was allowed to polymerize creating a barrier that prevented cellular attachment. TERT-pSUPER and TERT-siSFRP1 cells were plated in the wells and after 24 hours, FACs analysis was performed to compare the percentage of propidium iodide positive (FL2; dead cells) between cell lines. Scatter plot depicting FL2 positive cells vs. forward scatter (FSC-H) and oval represents area of viable cell not included in the analysis. Two separate experiments were performed in triplicate and bars represent mean ± SEM percent cell death. ***p < 0.001 (significantly different from control TERT-pSUPER cells using student's t-test) B, TERT-pSUPER and TERT-siSFRP1 cells were incubated with antibodies against human CD44-FITC and CD24-PE and labeled cells were analyzed by flow cytometry. FACS analysis reveals that TERT-pSUPER cells have a CD44^low/CD24^high phenotype and TERT-siSFRP1 cells exhibit a CD44^high/CD24^low phenotype.