a b c * * * P450arom mRNA Trascript Levels (Fold)

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a b c ** ** ** ** * * * - - - - P450arom mRNA Trascript Levels (Fold) Aromatase Activity (fmol/h/mg protein) Soft Agar Assay (colony #) G15 - - + shGPER - - + - AD - + - + MCF-7 G15 - - - MCF-7 TR2 + ANA - - + + MCF-7 MCF-7 TR2 Supplementary Fig. 1. a Aromatase (P450arom) mRNA content, evaluated by real-time RT–PCR, in MCF-7 and MCF-7 TR2 cells treated for 24 hours with vehicle (-) and/or the GPER antagonist G15 (1 mM). The values represent the means ± s.d. of three different experiments, each performed with triplicate samples. b Aromatase activity in MCF-7 and MCF-7 TR2 cells transfected with shGPER or treated or not for 24 hours with G15 (1 mM). The results obtained were expressed as fentomoles per hour and normalized to milligrams of protein (fmol/h/mg protein). c Soft-agar growth assay in MCF-7 TR2 cells treated with vehicle (-) or the aromatizable androst-4-ene-3,17-dione (AD, 10 nM) with or without the aromatase inhibitor anastrozole (ANA, 1µM). After 14 days of growth, colonies > 50 µm diameter were counted. Data are the mean colony number ± s.d. of three plates of three independent experiments. *p<0.05, **p<0.001.