FIGURE 4.

DEX-induced GR nuclear translocation and its phosphorylation at Ser-211 in MCF-7 cells treated with estrogen as detected by Western blot. A, cells were treated with hormones as described in Fig. 3. Nuclear and cytoplasmic protein samples were prepared using NE-PER nuclear and cytoplasmic extraction reagents and blotted with antibodies against Ser-211-phosphorylated GR. The membranes were stripped and reprobed with antibodies against total GR. Actin and TBP were used as loading controls for cytoplasmic and nuclear proteins, respectively. Images are representative of three independent experiments. Fold changes in the densitometry readings of nuclear total GR normalized to TBP (B) and Ser-211-phosphorylated nuclear GR normalized to total GR (C) in the cells treated with DEX alone (set as 1) versus cells cultured with E₂ and DEX are provided. NS, not significant.