unolocalized to some areas of the syncytiotrophoblast, leukocytes and to cohorts of EVT in the cell column but not to cytotrophoblasts. In the decidua, DPP1 localized to the glandular epithelium and to iEVT surrounding blood vessels. Glandular epithelial and iEVT localization was cytoplasmic, although in EVT the cytoplasmic localization was predominantly PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22180813 SU11274 site around the nucleus. Non-decidualized CM increased proDPP1 production. DPP1 could not be detected in EVT CM by western Annexin A2 Annexin A2 localized to iEVT in decidua basalis. Annexin A2 localized to the syncytiotrophoblast blotting. In EVT cell lysates 5 bands were visible; 52 kDa and 51 kDa, 34 kDa, 24 kDa and 7 kDa DPP1. By densitometry we found a significant increase in proDPP1 protein in EVT cell lysate following treatment with non-decidualized CM, but not decidualized CM compared to media control. There was no effect of HESC CM on mature or active DPP1. The mature cleavage form of DPP1 was only observed in EVT cell lysates from two of three women following treatment with HESC CM: in both women the mature cleavage form was present following treatment with non-decidualized CM and in only one woman it was found following treatment with decidualized CM. All three samples were from EVT isolated from weeks 78 of gestation. Lysosome associated membrane glycoprotein 1 LAMP1 localized to iEVT in decidua basalis. LAMP1 immunolocalized to the syncytiotrophoblast and leukocytes in the placental villous and to EVT in the cell column. In the decidua, LAMP1 localized to occasional cells in the glandular epithelium, HLAG negative cells surrounding the glandular epithelium and unremodelled blood vessels and iEVT surrounding remodelled blood vessels. LAMP1 could not be detected in EVT CM or cell lysates by western blotting. Discussion Here, for the first time, we identified proteins expressed by EVT in response to HESC-secreted factors. Our novel in vitro model identified proteins previously associated with diseases of pregnancy including preeclampsia suggesting our model can identify factors likely to be important in the pathogenesis of these diseases. Our Decidual Factors Alter Trophoblast Proteins 6 Decidual Factors Alter Trophoblast Proteins model also identified many proteins not previously known to be expressed by EVTs. We demonstrated that EVT expression of profilin 1, annexin A2 and proDPP1 was altered following treatment with HESC CM, suggesting that the degree of decidualization is critical for decidual-EVT crosstalk during EVT invasion. Further, our data suggests that non-decidualized factors may provoke EVTs to initiate a pro-inflammatory cascade, providing a potential mechanism for the pro-inflammatory state observed in preeclampsia. Recent studies have suggested that women with preeclampsia have impaired decidualization. 
Our identified proteins suggest that decidual regulation of EVT protein expression may be associated with the development of preeclampsia, however, future studies are required to validate the regulation of these proteins and their function. By fractionating our sample to identify proteins,30 kDa in size, we aimed to identify functional and regulatory proteins. The main functional groups of proteins identified by our proteomic approach were proteases/enzymes, membrane bound proteins, metal ion binding proteins, and proteins involved in redox regulation. Interestingly, treatment with decidualized CM was associated with an increase in the number of cell membrane protei
