Dmem f12 50 50 medium

The ovarian adenocarcinoma SKOV3ip1 and the ovarian immortalized INOF cell line were grown in DMEM/F12 (50/50) medium and OSE medium (Wisent), respectively. The medium in both cases was supplemented with 10% fetal bovine serum and 2 mM L-glutamine. Cell propagation and passaging were as recommended by ATCC (American Type Culture Collection). Cells were trypsinized and collected in 5 × 10⁶ pellets, resuspended in 700 µL TRIzol (Ambion) and kept at −80°C until RNA extraction.

All cell lines were cultured in antibiotic- and antimycotic-free media (Wisent). The ovarian adenocarcinoma SKOV3ip1 cell line was grown in DMEM/F12 (50/50) medium (Wisent), the breast adenocarcinoma MCF-7 cell line was grown in EMEM (Wisent) supplemented with 1 mM sodium pyruvate and MEM nonessential amino acid (Wisent), the immortalized normal ovarian fibroblast cell line INOF was grown in OSE (ovarian surface epithelium) medium (Wisent) and normal immortalized skin fibroblast BJ-Tielf was grown in alpha-MEM (Wisent). All media were supplemented with 10% fetal bovine serum and 2 mM L-glutamine. Cell propagation and passaging were as recommended by American Type Culture Collection. For transfection, cells were seeded at 350 000 cells per well in 6-well plates (BD Biosciences). Transfections were performed in suspension using Lipofectamine 2000 (Life technologies) according to the manufacturer's protocol and 20 nM of 21-mer siRNA (Sigma) for 48 h. All cell lines were actively growing when harvested. All knockdowns (KD) were done in duplicate using different siRNAs for each sample and were validated by qPCR (Supplementary File 1, Figure S1). The sequences of the siRNAs used are the following: CUUCUACCGUUCAGAUUCU (NOP58 KD 1), GACAAGUCCCAAACACAAA (NOP58 KD 2), GAUGGUCACACCAUAUGCA (RBFOX2 KD 1), CACCUCCGCAGAAUGGAAU (RBFOX2 KD 2).