Figure 2.

The GPR30–5′-flanking region is transactivated by EGF in SkBr3 and BT20 breast cancer cells. A–D, Cells were transfected with a reporter plasmid encoding the GPR30–5′-flanking region and treated with 100 nm E2, 1 μm G-1, 50 ng/ml IGF-I, or 50 ng/ml EGF, and 10 μm EGFR inhibitor tyrphostin AG 1478 (AG), 10 μm MEK inhibitor PD, 10 μm Src family tyrosine kinase inhibitor PP2, 10 μm PKA inhibitor H89, 10 μm PI3K inhibitor LY, as indicated. The luciferase activities were normalized to the internal transfection control and values of cells receiving vehicle (−) were set as 1-fold induction upon which the activity induced by treatments was calculated. Each data point represents the mean ± sd of three independent experiments performed in triplicate. ○, •, □, ▪, P < 0.05, for cells receiving vehicle (−) vs. treatment.