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

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. A) The relative expression of SLC22A23 under treatment with 5aza-dC increases in CESC as well as in OESC. In CESC, the increase in expression is dose dependent. B) The relative expression levels of OSR2 are decreased in CESC as well as in OESC under all tested doses of the demethylating agent. C) Treatment with 5aza-dC is associated with decreased relative expression levels of JAZF1 in both control and endometriotic stromal cells. D) Although in both CESC and OESC the ROR1 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. E) While the HAND2 gene relative expression levels decrease after demethylation treatment as shown in the left bar graph, the HAND2as levels change in the opposite direction; the expression levels are increased in both CESC and OESC, especially at the highest level of 5aza-dC. DAPK1 (F) and TGFBR1 (G) 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.