Figure 3
RhoB activation and regulation of estrogen receptor alpha expression with/without estradiol in MCF-7 cells. (A) Cells were deprived of estradiol (E2) for 3 days and then treated by E2 or epidermal growth factor (EGF) during the indicated times. GTP-bound RhoB and total lysates were collected. Protein expression was then analyzed. Representative of three to four independent experiments. (B) to (D) Cells were transfected with siControl (siC) or siB2, and 3 days later were deprived of E2 for 3 additional days and treated for 16 hours with E2 or ethanol. (B) Protein expression was then analyzed. Representative of three independent experiments. (C) Estrogen receptor alpha (ERα) expression was analyzed by immunocytochemistry. ImmunoReactive Score (IRS) shown in the upper-right corner. Representative of three independent experiments. (D) Expression of the ERα gene was then measured. Error bars represent the mean values ± standard deviation from triplicate conditions, representative of two independent experiments. Differences were considered statistically significant at *P < 0.01, Student's t test.