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nan12136-sup-0001-si.tif115K

Figure S1. ACTHR, CRHR1 and CRHR2 receptor genes are induced following ACTH and CRH treatment of primary adenoma cells. Following a 48 h. treatment of primary PA cells (5% CTS) with ACTH (2 × 10−9 M) or CRH (0.5 × 10−9 M) or Forskolin (4 × 10−5 M) the respective receptor gene expression was determined. Graph shows the fold significant increase above untreated control samples set to 1. Receptor genes are indicated and show the fold increase plus the standard error of the mean. For every drug induction a minimum of three experiments were performed for each PA. In the case of the ACTH PA 10 experiments for each hormone were performed.

nan12136-sup-0002-si.tif15628K

Figure S2. Localization and quantification of pCREB-Ser133 and Syncytin-1 to the same regions in PA and AH using IHC. Two PA (ACTH, Acro) IHC examples are shown from a total of six PA and one AH (pCREB-Ser133 and Syncytin-1). Quantitative analysis was performed with four 40-μm squared regions per tissue resulting in percentages of strong, intermediate, weak and negative pCREB-Ser133 nuclei expression correlated with Syncytin-1-positive cells in the same region.

nan12136-sup-0003-si.tif2989K

Figure S3. Cellular localization of TGF-β1 and TGF-β3 within PA and human term placental tissues. Examples of microscopic pictures examples of IHC tissue sections showing TGF-β1 and TGF-β3 expression with: CINon PA (top two panels) and ACTH PA (middle two panels). Note the cytoplasmic staining of both TGF-β1 and TGF-β3 throughout the PA. TGF-β1 and TGF-β3 has been determined to negatively regulate fusion of endometrial carcinoma cell fusion, placenta villous trophoblast cell fusion as well as osteoclast cell fusion [26,79,80]. The bottom two panels represent TGF-β1 and TGF-β3 localization in both human placental villi and the differentiated syncytiotrophoblast (SCT) (arrow) where cell fusion is reduced in the last trimester.

nan12136-sup-0004-si.docx20K

Table S1. Gene expression analyses of 20 different ERV env genes and the ERVW-5 gag gene in 92 PA and 8 normal AH. From left to right the gene name, control AH gene expression values (molecules/ng RNA) and standard error of the mean (SEM), PA gene expression values (molecules/ng RNA) and SEM with the P value indicated below each expression value. All significant values are indicated in bold. The total env molecules/ng RNA per patient cohort is indicated at the bottom above ERVW-5 gag.

nan12136-sup-0005-si.docx18K

Table S2. ERV gene stimulation of primary PA cell lines with ACTH, CRH and FK. From left to right columns represent primary cell type cultured including villous cytotrophoblasts (vCT); treatment of cells in 5% CTS with ACTH (2 × 10−9 M), CRH (0.5 × 10−9 M) or Forskolin (4 × 10−5 M); specific ERV genes are indicated with the fold increase plus the SEM above the untreated control. The nonparametric Mann–Whitney U test for independent samples was performed using IBM SPSS Statistics 21 for P value significances (*P < 0.05 and **P < 0.001).

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