Effect of Vav1 in cell cycle progression.

<p>(A) T47D cells were infected with lentivirus particles that expressed Vav1-specific shRNA or shRNA with a scramble sequence (served as a control). The homogenous cells were first synchronized to G0/G1 phase and then treated with DMSO as control or E₂ (10⁻⁷ mol/L) for 36 h before harvest. The expression of Vav1 and Cyclin D1 were analyzed by Western Blot with indicated antibodies respectively. The density of the bands was quantitated by Quantity One software (Bio-Rad, version 4.4.0, CA, USA). The bar chart represents the normalized protein level of Cyclin D1 to tubulin of three independent experiments. “**” indicates P<0.01 versus DMSO treatment of Control, and “a**” indicates P<0.01 versus E₂ treatment of Control by unpaired student T test. (B) Another aliquots of above cells were stained by PI and analyzed by flow cytometer for DNA contents. The decrease in the percentage of cells in G0/G1-phase before and after the treatment by DMSO or E₂ was calculated and plotted as y-axis and labeled on the bar graph. The differences between two T47D-Ctrl and T47D-ShVav1 cells were also marked. The data represented the mean ±S.D. of three independent experiments. “**” indicates P<0.01 versus corresponding control group by unpaired student T test.