E2-induced up-regulation of δEF1 is ER-α-dependent.

<p><b>A.</b> The ER-α-specific shRNA (shER-α) was transiently transfected into MCF-7 cells. Control cells (shControl) were treated with the scrambled shRNA. At the indicated time points, the expression of ER-α protein was verified by Western Blot. Actin was used to normalize the ER-α level. <b>B.</b> shER-α or shControl was transiently transfected into MCF-7 cells followed by treatment with 10⁻⁹ M E2. The expression of δEF1 mRNA was determined at the indicated time points after treatment using Q-PCR. GAPDH was used to normalize the δEF1 level. ** indicates p<0.01 in unpaired Student’s t-test compared with shControl. The data represent three independent experiments. <b>C.</b> shER-α or shControl was transiently transfected into MCF-7 cells followed by treatment with 10⁻⁹ M E2. The expression of δEF1 protein was determined at the indicated time points following treatment using Western Blot. Actin was used to normalize the δEF1 level. <b>D.</b> MCF-7 cells were pre-incubated with ICI 182,780 (1 µM) at the indicated time points. The expression of ER-α protein was verified by Western Blot. Actin was used to normalize the ER-α level. <b>E.</b> MCF-7 cells were pre-incubated with ICI 182,780 (1 µM) for 0.5 h followed by treatment with 10⁻⁹ M E2. The expression of δEF1 mRNA was determined at the indicated time points after treatment using Q-PCR. GAPDH was used to normalize the δEF1 level. ** indicates p<0.01 in unpaired Student’s t-test compared with control. The data represent three independent experiments. <b>F.</b> MCF-7 cells were pre-incubated with ICI 182,780 (1 µM) for 0.5 h followed by treatment with 10⁻⁹ M E2. The expression of δEF1 protein was determined at the indicated time points following treatment using Western Blot. Actin was used to normalize the δEF1 level.