Endocrine disruptors and human reproductive failure: the in vitro effect of phthalates on human luteal cells  Federica Romani, M.D., Anna Tropea, M.D., Ph.D., Elisa Scarinci, M.D., Alex Federico, M.D., Cinzia Dello Russo, M.D., Ph.D., Lucia Lisi, Ph.D., Stefania Catino, M.L.T., Antonio Lanzone, M.D., Rosanna Apa, M.D., Ph.D.  Fertility and Sterility  Volume 102, Issue 3, Pages 831-837 (September 2014) DOI: 10.1016/j.fertnstert.2014.05.041 Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Effect of di-(2-ethylhexyl) phthalate (DEHP), n-butyl benzyl phthalate (BBP), and di-n-butyl phthalate (DBP) on basal P4 release by human luteal cells. Luteal cells were cultured for 24 hours in medium alone (control), or with hGC (100 ng/mL), or DEHP (10–6, 10–7, 10–8, 10–9 M), or BBP (10–6, 10–7, 10–8, 10–9 M), or DBP (10–6, 10–7, 10–8, 10–9 M). Each value represents the mean ± SD of nine independent experiments, each done in triplicate. Results are expressed as a percentage of the control set equal to 100. ∗∗∗P<.001; ∗∗P<.01; ∗P<.05 vs. control values. Fertility and Sterility 2014 102, 831-837DOI: (10.1016/j.fertnstert.2014.05.041) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Effect of di-(2-ethylhexyl) phthalate (DEHP), n-butyl benzyl phthalate (BBP), and di-n-butyl phthalate (DBP) on P4-inducted hCG release by human luteal cells. Luteal cells were cultured for 24 hours in medium alone (control) or with hGC (100 ng/mL), or hGC (100 ng/mL) with DEHP added after 1 hour (10–6, 10–7, 10–8, 10–9 M), or BBP (10–6, 10–7, 10–8, 10–9 M), or DBP (10–6, 10–7, 10–8, 10–9 M). Each value represents the mean ± SD of nine independent experiments, each done in triplicate. Results are expressed as percentage of control set equal to 100. °°°P<.001 vs. C values; ∗∗∗P<.001 and ∗∗P<.01 vs. hCG values. Fertility and Sterility 2014 102, 831-837DOI: (10.1016/j.fertnstert.2014.05.041) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effect of di-(2-ethylhexyl) phthalate (DEHP), n-butyl benzyl phthalate (BBP), and di-n-butyl phthalate (DBP) on basal PGE2 release by human luteal cells. Luteal cells were cultured for 24 hours in medium alone (control) or with DEHP (10–6, 10–7, 10–8, 10–9 M), or BBP (10–6, 10–7, 10–8, 10–9 M), or DBP (10–6, 10–7, 10–8, 10–9 M). Each value represents the mean ± SD of seven independent experiments, each done in triplicate. Results are expressed as percentage of control set equal to 100. ∗∗∗P<.001; ∗∗P<.01; ∗P<.05 vs. control values. Fertility and Sterility 2014 102, 831-837DOI: (10.1016/j.fertnstert.2014.05.041) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Effect of di-(2-ethylhexyl) phthalate (DEHP), n-butyl benzyl phthalate (BBP), and di-n-butyl phthalate (DBP) on basal VEGF release by human luteal cells. Luteal cells were cultured for 24 hours in medium alone (control) or with CoCl2 (10 μM) or with DEHP (10–6, 10–7, 10–8, 10–9 M), or BBP (10–6, 10–7, 10–8, 10–9 M), or DBP (10–6, 10–7, 10–8, 10–9 M). Each value represents the mean ± SD of 10 independent experiments, each done in triplicate. Results are expressed as percentage of control set equal to 100. ∗∗∗P<.001; ∗∗P<.01 vs. control values. Fertility and Sterility 2014 102, 831-837DOI: (10.1016/j.fertnstert.2014.05.041) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions