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Figure 3 Expression of cancer stem cell markers in TNBC cells. Analyses were performed four days after ECP treatment. (A) Real-time PCR analysis of mRNA levels of pluripotency factors NANOG, OCT4 and SOX2 in MDA-MB-231 and SUM159-PT cells. RPLP0 mRNA was used to normalize variability in template loading. Data from control cells were set as 1, relative mRNA expression was evaluated by fold-change. (B,C) Flow cytometry analysis of cancer stem cell markers NANOG and CD49f in MDA-MB-231 cells. Cells were incubated with antibodies against NANOG and CD49f, and then analyzed by flow cytometry with FITC or APC filter, respectively. Isotype antibodies were used as controls. (B) Flow cytometry analysis of NANOGhigh population. Cells with fluorescence intensity ≥ log 102 were considered as NANOGhigh. Data were representative of 3 independent experiences (C) Flow cytometry analysis of CD49f positive sub-population. Quantitative graphics correspond to 3 independent experiments and illustrations are representative of 3 independent experiments. ns, not significant; *, p < 0.05; **, p < 0.01. Unpaired Student t-test.