RNA was extracted from snap frozen ovarian tumor sections, ascites-derived epithelial and mesenchymal cells and ovarian cancer cell lines stored in TRIzol® reagent (Ambion-Life Technologies, CA, USA) by the chloroform: phenol method as described previously (40 (link)). Five hundred ng of total RNA was reverse transcribed using the high-capacity cDNA Reverse Transcription Kit (Applied Biosystems, CA, USA), and both relative and absolute qRT-PCR amplification was performed using the Applied Biosystems ViiA 7 Real-Time PCR (Thermo Fisher Scientific, NSW, Australia) as described previously (40 (link)–42 (link)). Briefly, for absolute qRT-PCR method, samples were analysed against a known absolute quantity of the gene (fg), and these results were then standardized against an absolute quantity of 18S (fg) and this method was used for comparing tissue samples of different origin. While relative PCR compared Ct values relative to 18S Ct values (δΔCt) and was only used for samples within the same origin. All PCR reactions were performed in triplicate. The sequences and accession numbers of genes analyzed and primers used are listed in Supplementary Table 3
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