Fig. 2.

Effect of EE on nuclear content of phospho-ER-α (Ser118) in cultured hepatocytes. (A) Hepatocytes were incubated with EE (10 µM; 30, 60, or 120 minutes) or vehicle (C, control). (B) Hepatocytes were preincubated with ICI182/780 (1 µM; 30 minutes) and with EE (10 µM; 30, 60, or 120 minutes). Equal amounts of nuclear extract protein (15 µg) were loaded in all lanes. Uniformity of protein loading and transfer from gel to nitrocellulose membrane were controlled with Ponceau S and histone detection. Densitometric analyses were performed separately for phospho-ER-α and for total ER-α, and the phospho-ER-α/ER-α densitometry ratio was calculated and expressed as means ± S.D. (n = 4). Statistical analysis was performed using one-way analysis of variance, followed by Newman Keuls test. Values of P < 0.05 were considered to be statistically significant. a: significantly different from C, 60 minutes and 120 minutes, P < 0.05.