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The human colon cancer cell lines HCT116 was obtained from the American Type Culture Collection in 2001. Stocks were prepared after passage 2 and stored in liquid nitrogen. All experiments were performed with cells of passage of <8. These cell line was authenticated by morphologic inspection, short tandem repeat profiling, and Mycoplasma testing by the ATCC. Mycoplasma testing was done also by the authors. Cells were grown in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37°C in a humidified incubator with 5% CO2.The small interfering RNA (siRNA) for PTBP1 (Stealth Select RNAi, HSS143520) and negative control siRNA (Negative Control Hi GC) were purchased from Invitrogen. Cells were transfected with siRNA in 20 nmol/L concentration using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's protocols. |
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Protocol tips |
The human colon cancer cell lines HCT116 was obtained from the American Type Culture Collection in 2001. Stocks were prepared after passage 2 and stored in liquid nitrogen. All experiments were performed with cells of passage of <8. These cell line was authenticated by morphologic inspection, short tandem repeat profiling, and Mycoplasma testing by the ATCC. Mycoplasma testing was done also by the authors. Cells were grown in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37°C in a humidified incubator with 5% CO2.The small interfering RNA (siRNA) for PTBP1 (Stealth Select RNAi, HSS143520) and negative control siRNA (Negative Control Hi GC) were purchased from Invitrogen. Cells were transfected with siRNA in 20 nmol/L concentration using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's protocols. |
Publication protocol
The human colon cancer cell lines HCT116 was obtained from the American Type Culture Collection in 2001. Stocks were prepared after passage 2 and stored in liquid nitrogen. All experiments were performed with cells of passage of <8. These cell line was authenticated by morphologic inspection, short tandem repeat profiling, and Mycoplasma testing by the ATCC. Mycoplasma testing was done also by the authors. Cells were grown in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37°C in a humidified incubator with 5% CO2.The small interfering RNA (siRNA) for PTBP1 (Stealth Select RNAi, HSS143520) and negative control siRNA (Negative Control Hi GC) were purchased from Invitrogen. Cells were transfected with siRNA in 20 nmol/L concentration using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's protocols
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