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MB Sample ID: SA322337
Local Sample ID: | NEG_EV1_1 |
Subject ID: | SU003072 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
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Treatment:
Treatment ID: | TR003081 |
Treatment Summary: | shRNAs in the pLKO-puro lentiviral vector, were packaged in Lenti-X 293T Cells (Takara Bio, Cat. 632180) by transient transfection, in combination with the packaging constructs psPax2 (Addgene, Cat. 12260) and pMD2.G (Addgene, Cat. 12259). Transfections were carried out using the Lipofectamine 3000 Transfection Reagent (Thermo Fisher Scientific, Cat. L3000015) and the Opti-MEM Reduced Serum Medium (Fisher Scientific, Cat. 31–985-070), according to the manufacturer’s protocol. The supernatants were collected 48h and 72h after the transfection. MDA-MB-231 and MDA-MB-468 cells were infected with the viral supernatants, in the presence of 8 μg/mL polybrene (Millipore-Sigma, Cat. 107689). Infected cells were selected with puromycin for 48h (Gibco, Cat. A11138) (10 μg/mL). |