Fig. 5

Relationship between PLC-β2 and MDA-MB-231 sub-populations with a stem-like phenotype. In a MDA-MB-231 cells transfected with siRNAs specific for PLC-β2 (PLC-β2 siRNAs) or with a construct expressing human PLC-β2 (Over PLC-β2) were subjected to a bi-parametric flow cytometry by direct staining with the anti-CD133 and anti-EpCAM fluorescent antibodies to evaluate the number of CD133⁻/EpCAM⁻, CD133⁻/EpCAM⁺, CD133⁺/EpCAM⁻ and CD133⁺/EpCAM⁺ cells. Non-silencing scramble siRNAs or an empty vector were used as control (Ctrl). The data, representative of three separate experiments, are presented on pie charts and the percentage of each cellular subset is reported. In b MDA-MB-231 cells in which PLC-β2 was forcedly silenced (PLC-β2 siRNAs) or over-expressed (Over PLC-β2) were simultaneously stained with APC-conjugated anti-CD44, FITC-conjugated anti-EpCAM and PE-conjugated anti-CD133 and subjected to flow cytometry evaluation of CD44, EpCAM and CD133 surface levels. Data are the mean of three separate experiments performed in triplicate ± SD. *P < 0.05