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. 2016 Dec 24;156(1):54–71. doi: 10.1093/toxsci/kfw234

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© The Author 2016. Published by Oxford University Press on behalf of the Society of Toxicology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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FIG. 3 — Extracts of soil samples in close proximity to a landfill site contain a chemical(s) that activate the mouse ERα in a murine ductal (LTPA) cell line. A, Western Blot (20 μg total protein/lane) for the expression of the mouse ERα protein in LTPA and HEK293 cells transfected with the expression construct encoding the mouse ERα cDNA sequence. B, immunocytochemical analysis for ERα expression in LTPA or HEK293 cells transfected with an expression vector encoding the mERα cDNA sequence. C, luciferase reporter gene (3XERE-TATA) assay in LTPA cells co-transfected with expression constructs encoding the indicated protein. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus the equivalent expression vector transfected DMSO vehicle-treated cells; #significantly different (P < .05) versus equivalent treatments in the absence of ICI182780-treated cells using the Student’s t-test (2-tailed). D, luciferase reporter gene (3XERE-TATA) assay in LTPA cells co-transfected with expression construct encoding the mERα and comparison to cells transfected with an empty expression construct. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus empty vector transfected cells treated with the same concentration of E2 or EE. E, luciferase reporter gene (3xERE-TATA) assay in LTPA cells. Cells were transfected as outlined in the methods section and treated with 0.1% v/v ethanol extracts, E2 or EE for 24 h prior to analysis. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus the equivalent expression vector transfected DMSO vehicle-treated cells; #significantly different (P < .05) versus equivalent treatments in the absence of ICI182780-treated cells using the Student’s t-test (2-tailed).

FIG. 3 — Extracts of soil samples in close proximity to a landfill site contain a chemical(s) that activate the mouse ERα in a murine ductal (LTPA) cell line. A, Western Blot (20 μg total protein/lane) for the expression of the mouse ERα protein in LTPA and HEK293 cells transfected with the expression construct encoding the mouse ERα cDNA sequence. B, immunocytochemical analysis for ERα expression in LTPA or HEK293 cells transfected with an expression vector encoding the mERα cDNA sequence. C, luciferase reporter gene (3XERE-TATA) assay in LTPA cells co-transfected with expression constructs encoding the indicated protein. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus the equivalent expression vector transfected DMSO vehicle-treated cells; #significantly different (P < .05) versus equivalent treatments in the absence of ICI182780-treated cells using the Student’s t-test (2-tailed). D, luciferase reporter gene (3XERE-TATA) assay in LTPA cells co-transfected with expression construct encoding the mERα and comparison to cells transfected with an empty expression construct. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus empty vector transfected cells treated with the same concentration of E2 or EE. E, luciferase reporter gene (3xERE-TATA) assay in LTPA cells. Cells were transfected as outlined in the methods section and treated with 0.1% v/v ethanol extracts, E2 or EE for 24 h prior to analysis. Data are the mean and SD normalized luciferase activity from 3 entirely separate determinations of the same conditions performed within the same experiment, typical of at least 3 separate experiments performed at different times. *Significantly different (P < .05) versus the equivalent expression vector transfected DMSO vehicle-treated cells; #significantly different (P < .05) versus equivalent treatments in the absence of ICI182780-treated cells using the Student’s t-test (2-tailed).