Fig. 3. The over-expression of ETV7 influences the expression of breast cancer stem cell markers and mammosphere formation efficiency.

A, B CD44-APC and CD24-FITC staining and flow cytometry analysis in MCF7 (A) and T47D (B) Empty and ETV7 cells. On the left, a representative dotplot of the results obtained at FACS Canto A; the histogram on the right summarizes the percentage of CD44⁺/CD24⁻ cells in Empty and ETV7 over-expressing cells. C Western blot analysis of EpCAM expression in MCF7 Empty and MCF7 ETV7 cells. The correct band is indicated with an arrow. HSP70 was used as a loading control. On the right of each blot is indicated the approximate observed molecular weight. D A representative image of first-generation mammospheres obtained from MCF7 Empty and MCF7 ETV7 cells, respectively. The scale bar is indicated. E The percentage of mammosphere formation efficiency (% MFE) in MCF7 Empty and ETV7 calculated as the number of mammospheres per well/number of cells seeded per well × 100. F % MFE in second-, third-, and fourth-generation mammospheres obtained by passaging the mammospheres every 7 days. Bars represent the averages and standard deviations of at least three independent experiments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001.