ute wortmanin pre-treatment. Similar downregulation of Dnmt3A and Dnmt3L gene expression was observed in NT2/D1 cells. Hence, M4- and PI3K-dependent repression of DNMT3B and DNMT3L was further observed at the protein level. This suggests that the ERa36-dependent M4 signaling ending at target genes involved in DNA-methylation status could be a common feature of both seminoma and embryonal carcinoma cells. Discussion Although numerous chemicals are now known or suspected to have endocrine disruption effects, a relevant classification based on comprehensive understanding of their mode of action and targets is still failing. More confusing is the wide variety of cocktails detected in the environment, when trying to decipher dose response consequences for lifelong human exposure. In the present study, we chose to focus on a well defined mix of alkyphenols – M4 -composed of 4-tert-octylphenol and 4-nonylphenol. Despite the burden of recent research on BPA which belongs to the same chemical 17149874 family and exert MedChemExpress 2883-98-9 various estrogenic effects, 4tert-octylphenol and 4-nonylphenol are still 
neglected. High doses of tert-octylphenol or nonylphenol ranging from 25 to 200 mg/kg bw were previously shown to significantly decreased sperm count and quality in male mice, and affect uterine weight, vaginal opening and reproductive ability in female rats. However, both molecules have never been associated in a realistic mixture Alkylphenols Trigger ERalpha36 Mitogenic Signaling mimicking daily human contamination from household products, cosmetics and food. Here, estrogen-like mechanisms of action were addressed in a model of TGCT lacking the long form of ERa receptor. As in the case of 17b-estradiol or its BSAcoupled counterpart, M4 doses ranging from 10 nM to 0.1 nM stimulated both seminoma and embryonal carcinoma cell pro- liferation in a non monotonic dose-response manner. Alkylphenols Trigger ERalpha36 Mitogenic Signaling respond to M4 in a non-monotonic way, as observed for in vitro cell proliferation or that a mild toxicity could appear after exposure to high 10712926 doses of the mix. Taken together, these results suggest that alkylphenol exposure may on the one hand, alter normal germ cell multiplication and differentiation during development through mutagenic or clastogenic mechanisms at high doses as described by others and on the other hand, elicit neoplastic germ cell proliferation at low doses, as shown in this study. Therefore we investigated the rapid non-genomic transduction pathways potentially involved. Whereas estradiol and BPA were previously shown to bind and exert such mitogenic effects through GPER in both SKBR3 breast cancer cells and JKT-1 seminoma derived cells, we demonstrated that M4 acts mainly via an ERa36-dependent pathway. Indeed, we evidence here that M4 triggers PI3K activity and CREB phosphorylation. Nevertheless, preliminary data indicate that both GPER and ERa36 may activate downstream signaling such as src phosphorylation and thus modulate the expression of M4 target gene subclasses as well as cell proliferation. Since GPER was shown to partially govern ERa36 expression in our TCam-2 model and may collaborate with ERa36 for estrogenic activities in other cancer cell lines, it would be relevant to test the participation of ERa36 in alkylphenol response in hormone-sensitive cancers such breast or prostate cancers. The microarray analysis performed in order to describe the gene expression pattern of M4-treated TCam-2 cells, indicated th
