Supplementary Figures 1 - 6 from SERMs Attenuate Estrogen-Induced Malignant Transformation of Human Mammary Epithelial Cells by Upregulating Detoxification of Oxidative Metabolites

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PDF file - 391K, Supplementary Figure S1: Recombinant COMT (10 microg/mL) was incubated with S-(5'-adenosyl)-L-methionine (500 nM) and raloxifene, DMA, or bazadoxifene (1 microM ) at 37 degrees C for 5 minutes prior to the addition of 6,7-dihydroxycoumarin (5 microM). Supplementary Figure S2: Chromatograms showing (A) OHE2 / E1 metabolites and (B) MeOE2 / E1 metabolites in E2 (1 microM) treated MCF-10A cell samples. Relatively higher amounts of E2 metabolites were detected compared to E1 metabolites in E2 treated cell samples. Supplementary Figure S3: Effect of SERMs on CYP450 1B1 and 1A1 enzyme expression. MCF-10A cells were treated with E2 (1 microM) in the presence and absence of SERMs (1 microM). Supplementary Figure S4: Inhibition of CYP450 1B1 by SERMs. Recombinant CYP450 1B1 was incubated in potassium phosphate buffer (50 mM, pH = 7.4) in the presence of NADPH (1 mM) and different concentrations of SERMs (0, 10, 20, 30, 40, 50, 60 microM). Supplementary Figure S5: Representative western blot images showing the (A) effect of SERMs on GST-P1,catechol-O-methyl transferase (COMT) and (B) SULT 1A1, SULT1E1 and NQO1 in MCF-10A cells. Cells were treated with E2 (1 microM) in the presence and absence of SERMs (1 microM) for 6 days. Supplementary Figure S6: MCF-7 cells were were plated (4 x 10^5 cells) in 12 well plates.

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