1. Aryl hydrocarbon receptor antagonists attenuate the deleterious effects of benzo[a]pyrene on isolated rat follicle development 
Michael S. Neal, Anne M. Mulligan Tuttle, Robert F. Casper, Alex Lagunov, Warren G. Foster 
Reproductive BioMedicine Online 
Volume 21, Issue 1, Pages (July 2010)
DOI: /j.rbmo
Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

2. Figure 1
FSH-stimulated growth of preantral isolated rat follicles in vitro. Increasing concentrations of benzo[a]pyrene (black bar) attenuated growth and reached statistical significance at a concentration of 1.5ng/ml. Co-treatment with 5μmol/l resveratrol (grey bar) ameliorated the deleterious effect of benzo[a]pyrene. Results are expressed as mean±SEM of six replicates of 96 follicles per culture. ∗P<0.05.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

3. Figure 2
FSH-stimulated growth of preantral isolated rat follicles in vitro. Increasing concentrations of benzo[a]pyrene (black bar) attenuated growth and reached statistical significance at a concentration of 1.5ng/ml. Co-treatment with 5μmol/l 3′,4′-dimethoxyflavone (grey bar) ameliorated the deleterious effect of benzo[a]pyrene. Results are expressed as mean±SEM of seven replicates of 96 follicles per culture. ∗P<0.05.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

4. Figure 3
Representative images of isolated rat follicles after 5days of culture in: (a) follicle culture media (FCM) alone (control); (b) FCM supplemented with 1.5ng/ml benzo[a]pyrene; or (c) FCM supplemented with 1.5ng/ml benzo[a]pyrene co-incubated with an aryl hydrocarbon receptor antagonist (resveratrol). Arrows point to the oocyte surrounded by granulosa cells, which are healthier in appearance and growth in both a and c compared with b. Bars=20μm.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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5. Figure 4
Isolated rat follicle cell viability, measured with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, for follicles exposed to varying concentrations of benzo[a]pyrene, alone (black bar), with 5μmol/l resveratrol (white bar) or 5μmol/l 3′,4′-dimethoxyflavone (grey bar). Values are expressed as percentage of controls. Follicles exposed to concentrations of 5.0ng/ml benzo[a]pyrene or greater had significantly lower cell viability compared with controls and follicles co-incubated with resveratrol or 3′,4′-dimethoxyflavone (t-test). ∗P<0.001.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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6. Figure 5
Concentrations of oestradiol (pg/ml) in spent media from the isolated follicle culture assay. Oestradiol concentration was inversely related to increasing concentration of benzo[a]pyrene (black bar), decreased in a dose-dependent manner and was significantly lower than controls (∗). Co-treatment with 5μmol/l 3′,4′-dimethoxyflavone (grey bar) resulted in a higher oestradiol concentration but still lower than controls. At a concentration of 50.0ng/ml, there was a significant difference in oestradiol concentration between follicles grown in benzo[a]pyrene alone compared with those co-incubated with 3′,4′-dimethoxyflavone (∗∗). ∗P<0.05; ∗∗P<0.05).
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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7. Figure 6
Concentrations of anti-Müllerian hormone (AMH) measured by enzyme-linked immunosorbent assay in the spent media of isolated rat follicle cultures exposed to increasing concentrations of benzo[a]pyrene alone (black bar) or co-incubated with 5μmol/l of 3′,4′-dimethoxyflavone (white bar). Values are expressed as percentage of controls. AMH concentrations were significantly lower in follicles exposed to increasing concentrations of benzo[a]pyrene. AMH concentrations were similar in spent media samples from follicles co-cultured with 3′,4′-dimethoxyflavone and were significantly different than their treated counterparts. ∗P<0.001; ∗∗P<0.05.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions