Validation of RNA-sequencing by Q-PCR and effects of 5Aza-dC on a subset of DM and DE genes in CESC and OESC.

<p>This figure shows average gene expression levels in CESC and OESC, from two or three biological replicates, after treatment with 5aza-dC for 72 hrs. All values were normalized to the mean of the non-treated CESC controls (0) set to 1. <b>A)</b> The relative expression of <i>SLC22A23</i> under treatment with 5aza-dC increases in CESC as well as in OESC. In CESC, the increase in expression is dose dependent. <b>B)</b> The relative expression levels of <i>OSR2</i> are decreased in CESC as well as in OESC under all tested doses of the demethylating agent. <b>C)</b> Treatment with 5aza-dC is associated with decreased relative expression levels of <i>JAZF1</i> in both control and endometriotic stromal cells. <b>D)</b> Although in both CESC and OESC the <i>ROR1</i> expression levels decrease under 5aza-dC treatment, in CESC the effect on gene expression is dose dependent and more pronounced compared to the endometriotic cells. <b>E)</b> While the <i>HAND2</i> gene relative expression levels decrease after demethylation treatment as shown in the left bar graph, the <i>HAND2as</i> levels change in the opposite direction; the expression levels are increased in both CESC and OESC, especially at the highest level of 5aza-dC. <i>DAPK1</i> <b>(F)</b> and <i>TGFBR1</i> <b>(G)</b> expression levels are not influenced by the drug in CESC or in both CESC and OESCs, respectively. For direct comparison of Q-PCR and RNA-seq data, the differences in expression between CESC and OESC, identified by RNA-seq method are indicated below each graph. CESC, control endometrial stroma cells, OESC, ovarian ectopic endometriosis stroma cells. In each graph, † indicates conditions with biological duplicates.