Xfect transfection agent

Human ovarian cancer cell lines HO8910 was obtained from department of pathophysiology of Second Military Medical University. SKOV3 and its cognate Cisplatin-resistant cells SKOV3/DDP were obtained from ATCC (American Tissue Culture Collection). SKOV3.ip cells were provided by the Department of Gynecology and Obstetrics, Shanghai First People's Hospital (obtained from ATCC). Cells were maintained in RPMI-1640 medium with 10% fetal bovine serum at 37°C in a humidified 5% CO₂ atmosphere. Cisplatin, doxycycline, SDF-1α, antibody against β-actin were purchased from Sigma-Aldrich. Antibodies against CXCR4, phosphor-Akt, total Akt, phosphor-ERK, total ERK, MMP-2, MMP-9, caspase-3 were purchased from Cell Signaling Technology. Transwell kit was purchased from BD, USA. ELISA kit was purchased from R&D, USA. Cell apoptotic kit was purchased from Roche, USA. Small interfering RNAs were synthesized by Genepharma Company (Shanghai, China). Xfect Transfection Agent was purchased from Clontech, USA. RNeasy minikit was purchased from QIAGEN, Clifton Hill, Australia.

The vector used was taken from Addgen (http://www.addgene.org/) (Plasmid 26817): pcDNA3.3_SOX2; and transformation into SW1736 cells was performed using Xfect transfection agent (Clontech Laboratories, Inc. A Takara Bio Company) according to the manufacturer's instructions. The transfection agent and plasmid were added to the cells and incubated for 72 hours for RNA extraction and analysis.