Additionally, BMP- and WNT signaling-associated molecules and as well as pluripoteny-related gene were upregulated 8 days after NOGGIN treatment (Fig 4C)

Additionally, BMP- and WNT signaling-associated molecules and as well as pluripoteny-related gene were upregulated 8 days after NOGGIN treatment (Fig 4C). unmethylated CpGs and filled circles methylated CpGs.(TIF) pgen.1005415.s004.tif (7.7M) GUID:?FFF38186-7C21-4734-8CA7-EBCFCAB629D2 S5 Fig: Interactive network prediction and verification of Gex data in EC cell lines. (A, B) STRING-based conversation prediction of genes commonly upregulated in TCam-2 1 (A) and 6 (B) weeks after xenografting. (C) qRT-PCR analysis of indicated seminoma and EC markers in parental TCam-2 and EC cell lines (2101EP, NCCIT, NT2/D1, 833KE, H12).(TIF) pgen.1005415.s005.tif (656K) GUID:?405B8CC6-4520-4789-BEAC-A14541D4922E S6 Fig: Signaling pathways that drive reprogramming of TCam-2 cells into an EC. (A) cDNA microarray expression data of indicated signaling pathway-associated genes during SET and in 2102EP control cells. (B) Western blot analysis of SOX2 and pSMAD1 /5 expression in in vitro cultivated and xenografted TCam-2 cells (4 weeks). (C, D) Western blot analysis of SMAD1 /5-phosphorylation in TCam-2 treated with NOGGIN (C) and LDN193189 (D) or corresponding solvents. (E) qRT-PCR analysis of indicated genes in TCam-2 cells treated with the BMP inhibitor LDN193189 for 48C96 h. (F) Western blot analysis of SMAD2 /3-phosphorylation 72 h after treatment of TCam-2 cells with recombinant NODAL or the solvent. (G) qRT-PCR analysis of indicated genes in TCam-2 cells treated for 24 and 72 h with recombinant NODAL.(TIF) pgen.1005415.s006.tif (483K) GUID:?868DA9C6-A89D-4DBB-9740-478152F4F0B5 S7 Fig: GCC-TMA-IHCs and western blot analysis of ZIC3 expression. (A) Examples of ID1, TFAP2C, SOX2 and beta-CATENIN IHC in CIS, seminoma and EC tissues. Scale bars: 100 m. (B) Western blot analysis of ZIC3 and SOX2 expression in indicated GCC cell lines CI 972 and human fibroblasts. (C) XY-diagrams illustrating the correlation /reciprocal correlation of to and in GCC tissues.(TIF) pgen.1005415.s007.tif (5.8M) GUID:?17A2C537-398E-4992-BA3A-B573A81E2630 S1 Data: Summarized cDNA expression and DNA methylation results. (XLSX) pgen.1005415.s008.xlsx (2.9M) GUID:?B2F14CC4-FBE1-4804-BD05-B5E1227A98C7 S1 Table: Antibodies used in this study. (XLSX) pgen.1005415.s009.xlsx (60K) GUID:?34367796-0D3F-4664-8FC4-07A5F7104B92 S2 Table: Oligonucleotides used in this study. (XLSX) pgen.1005415.s010.xlsx (58K) GUID:?270FEF32-9C96-4CDE-87F1-8909F7A8AB0D Data Availability StatementAll relevant data are within the paper and its Supporting Information files except for the microarray data. The microarray data sets are publicly available via GEO ( (GSE60698, GSE60787). Abstract Type II germ cell cancers (GCC) can be subdivided into seminomas and non-seminomas. Seminomas are similar to carcinoma in situ (CIS) cells, the common precursor of type II GCCs, with regard to epigenetics and expression, while embryonal carcinomas (EC) are totipotent Klf4 and differentiate into teratomas, yolk-sac tumors and choriocarcinomas. GCCs can present as seminomas with a non-seminoma component, raising the question if a CIS gives rise to seminomas and ECs at the same time or whether seminomas can be reprogrammed to ECs. In this study, we utilized the seminoma cell line TCam-2 that acquires an EC-like status after CI 972 xenografting into the murine flank as a model for a seminoma to EC transition and screened for factors initiating and driving this process. Analysis of expression and DNA methylation dynamics during transition of CI 972 TCam-2 revealed that many pluripotency- and reprogramming-associated genes were upregulated while seminoma-markers were downregulated. Changes in expression level of 53 genes inversely correlated to changes in DNA methylation. Interestingly, after xenotransplantation 6 genes (in murine and human ESCs induced is normally repressed by [17]. Further, it is known that SOX17 antagonizes WNT signaling, which has been suggested to demarcate seminomas from ECs [18] CI 972 [19] [20]. Signaling pathways in normal and malignant germ cells The members of the TGF-beta superfamily.