Figure 3. HER2 overexpression and activity affect hMena/hMena^11a expression in human breast cancer cell lines.

A. HER2 transfection affects hMena expression in MCF7 breast cancer cells. Western blot analysis of MCF7-pcDNA3 and MCF7-HER2 lysates (50 µg) with the indicated antibodies. Two different antibodies for hMena have been used: the pan-hMena antibody, recognizing hMena and hMena^11a as a doublet of 88–90 kDa, and the anti-hMena^11a specific antibody. Membranes were sequentially stripped and reprobed with the indicated total and phospho-specific antibodies. B. hMena/hMena^11a expression in HER2 overexpressing breast cancer cell lines. Western blot analysis of BT474 and SKBr3 breast cancer cell line lysates (50 µg) with the indicated antibodies. As loading control, blots were probed with anti-Actin mAb (1 µg/ml). C. HER2 transfection, NRG and EGF treatment affect hMena expression at mRNA level. QRT-PCR analysis of hMena and housekeeping β-Actin mRNA levels performed on 10 ng of total RNA from the untreated MCF7-pcDNA3, HER2 transfected MCF7, or NRG1 treated MCF7 breast cancer cell line and SKBr3 cells either untreated or treated with EGF. Results are given as the ratio between hMena and β-Actin genes relative to the internal control.