Fig 1.
Chemical structures and identification numbers for compounds analyzed.
Each compound is identified by an abbreviation, full name, and a PubChem compound identification (or CAS) number.
Fig 2.
PRL-HeLa analysis of estrogenic and anti-estrogenic activity of investigational chemical compounds on ERα.
(A) Experimental workflow for analysis of investigational molecules. Percentage of cells with an array in GFP-ERα:PRL-HeLa cells after 1 hour of treatment with maximal dose of compound alone or with 10 nM E2 after 1 hour (B) or 24 hours (C) of exposure. Antagonism experiments were done combining 10nM E2 with 5 μM of the compounds of interest. (D) Dose-response curves of percent of cells with arrays for E2, BPA and the seven investigational compounds. (E) Effects on SRC-3 coactivator and RNA polymerase II levels at visible arrays after 1 hour of treatment with 10 nM E2 in the presence of maximal dose of compound. (F) Effects on de novo mRNA production at the PRL array after 1 hour of treatment with compound alone or with 10 nM E2. (G) Effects on GFP-ERα expression level after 24 hours of treatment with maximal dose of compound. All data represent mean of a minimum of 4 experiments ± standard deviation. (*) or (**) indicates a response significantly (p < 0.05) above or below control treatment.
Fig 3.
PRL-HeLa analysis of ERβ estrogenic and anti-estrogenic activity of investigational chemical compounds.
Percentage of cells with an array in GFP-ERβ:PRL-HeLa cells after 1 hour of treatment with maximal dose of compound alone or with 10 nM E2 after 1 hour (A) or 24 hours (B) of exposure. (C) Dose-response curves of percent of cells with arrays for E2, BPA and the seven investigational compounds (D) Effects on GFP-ERβ expression level after 24 hours of treatment with maximal dose of compound. All data represent mean of a minimum of 4 experiments ± standard deviation. (*) or (**) indicates a response signifcantly (p < 0.05) above or below control treatment.
Fig 4.
Chemical compound effect on an integrated ERE-driven luciferase reporter in the ER responsive MCF-7 cell line.
MCF-7-MAR-ERE-Luc were treated with indicated compounds at 5 μM, 5 μM + 10 nM E2, or 2 μM + 10 nM E2 for 24 hours before total luciferase activity determined. The %E2 calculated using control wells treated with E2 (10 nM) or DMSO. Dashed line indicates 10 nM E2 control response. All data represent mean of 3 experiments ± standard deviation. (*) or (**) indicates a response significantly (p < 0.05) above or below control treatment.
Fig 5.
Chemical compound effect on an integrated ARE-containing probasin-mCherry-NLS reporter in the AR responsive LNCaP cell line.
The nuclear accumulation of mCherry fluorescent protein in 2PB-mCherry-NLS:LNCaP cells treated with indicated compounds either alone at concentrations from 50 pM to 5 μM (A) or at 5μM with DHT at ranging from 0.1 nM to 30 nM (B) for 48 hours. Compounds that induced either a significant androgen-like or anti-androgen-like response in the presence of DHT are indicated. All data represent mean of 3 experiments ± standard deviation.
Fig 6.
Clustering analysis of investigational chemical compounds and previously studied bisphenol analogs activity across multiple in vitro assays.
Data in each assay row were range normalized before clustering analysis. Clustering was performed using Euclidean distance for both the compounds and the assays.