Figure 3:

Effects of genistein on ERα and ERβ expression levels in human uterine LM cells and human uterine SMC.

(A) Real-time RT–PCR analysis of the relative expression of ERα and ERβ mRNA in LM cells and SMC in 0.3% DMSO (Control) or treated with 1 µg/ml of genistein (Gen) in phenol red-free DMEM/F-12 containing 10% charcoal-stripped fetal calf serum for 24 h. HPRT served as a loading control. Experiments were repeated at least three independent times with duplicate samples. Fold changes were calculated by dividing values on the y-axis with the vehicle control (set at 1). Results are presented as mean ± SEM (n = 3). (B) A representative western blot of the protein expression of ERα and ERβ in LM cells and SMC cultured in phenol red-free DMEM/F-12 containing 10% charcoal-stripped FBS for 24 h. Molecular weight markers are indicated on the right of the blot. (C) Time course of the protein expression of ERα and ERβ in LM cells in 0.3% DMSO (Control) at 0 min or treated with 1 µg/ml of genistein in phenol red-free DMEM/F-12 containing 10% charcoal-stripped fetal calf serum for 5, 10, 15, 30, 60 min, 24, 72 and 168 h. HPRT was served as a loading control. (D) Time course of the protein expression of ERα and ERβ in SMC treated with 0.3% DMSO (Control) at 0 min or 1 µg/ml of genistein in phenol red-free DMEM/F-12 containing 10% charcoal-stripped fetal calf serum for 5, 10, 15, 30, 60 min, 24, 72 and 168 h. The blots presented are representative examples of experiments that were performed at least three times with repetitive results. Molecular weight markers are indicated on the right of the blot. The histograms shown below are the quantitative representation after densitometry of data (mean ± SD) of three independent experiments. Values of ERα and ERβ band densities in LM cells and SMC were divided by the vehicle control (set at 1).*P < 0.05 versus vehicle control containing 0.3% DMSO. **P < 0.05 compared to 24 h.