PRJNA530710
Expression data of  melanoma cell lines after SIRT2 depletion
Scope: to analyze SIRT2-dependent gene expression in melanoma cells of vertical growth and metastasis.1. The human melanoma cell line WM853 (stage: primary/vertical growth) was purchased from Rockland (Rockland, Limerick, PA, USA) and was maintained in Tumor Specialized Medium containing 80% MCDB153 (Sigma-Aldrich), 20% Leibovitz's (Sigma-Aldrich), 2% fetal bovine serum (Pan Biotech GmbH), 1.68 mM calcium chloride, and 1.2 g (11.2 mM) sodium bicarbonate. WM853 cells were transfected with short hairpin RNA (shRNA) plasmids: scrambled negative control non-effective shRNA (TR30012) or SIRT2 (T301692D) (Origene Tech.). Transfection was performed with Metafectene PRO (Biontex) following the manufacturer's instructions. The cells were then cultured by replacing the medium every three days with complete medium containing 0.4 µg/ml puromycin for one month, during which stable clones were selected: WM853 SCW3 (control) and WM853 SSW30 ( with downregulated SIRT2). RNA was isolated from SCW3 and SSW30 cells (4 replicates per each cell line).2. The human melanoma cell line MDA-MB-435S (stage: metastatic) was obtained from the ATCC and was maintained in Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum. MDA-MB-435S cells were transfected with short hairpin RNA (shRNA) plasmids: scrambled negative control non-effective shRNA (TR30012) or SIRT2 (T301692D) (Origene Tech.). Transfection was performed with Metafectene PRO (Biontex) following the manufacturer's instructions. The cells were then cultured by replacing the medium every three days with complete medium containing 1 µg/ml puromycin for one month, during which stable clones were selected: MDA-MB-435S SCM1 (control) and MDA-MB-435S SSM15 (with downregulated SIRT2). RNA was isolated from SCM1 and SSM15 cells (4 replicates per each cell line).