Figure 2

Vav3 is involved in growth of breast cancer cells. (A) Expression analysis of Vav3 and ERα in breast cancer MCF7 and T47D cells and nontumoral breast epithelial MCF-10A cells. The cell extracts were prepared from T47D, MCF7, and MCF-10A cells and subjected to western blot analysis for Vav3 and ERα. β-actin was served as loading control. (B) MCF7 cells were transiently transfected with Vav3 expression vector or control empty vector and then cultured in stripped medium in the absence or presence of EGF for 5 days, followed by MTT assay. The data was presented as absorbance at OD 570 nM. (C) MCF7 cells were transiently transfected with Vav3 expression vector or empty pHEF vector for 3 days, followed by cell extracts preparation and western blot analysis for Vav3. β-actin was served as loading control. (D) Knock down expression of Vav3 upon transfection of Vav3 siRNA. T47D cells were transfected with 5 pM/well of siVav3-247 or control-247 in 6-well plate for 3 days. The cell extracts were prepared and subjected to western blot analysis for Vav3. β-actin was served as loading control. (E and F) MCF7 cells and T47D cells (2500 cells/well in 96-well plate) were transiently transfected with siVav3-247 or control-247 at the concentrations of 0, 0.15, 0.3, 0.6, 1.2, 2.5, 5.0 pM/well using Lipofectamine 2000 for overnight. Then, the cells were cultured in stripped medium without or with E2 (10^-9 M) for 5 days, followed by MTT assay. The data was presented as absorbance at OD 570 nM.