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siRNA concentration-20 nM
ICAM-1 siRNA: 5′-CGGCUGGAGCUGUUUGAGATT-3′ and 5′-GGAGUGAUUUUU CUAUCGGTT-3′
Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).
Incubate for 5 minutes at room temperature and add to cells.
Replace medium after 5h and re- incubate cells for 2 days at 37°C. |
|
Protocol tips |
siRNA concentration-20 nM
ICAM-1 siRNA: 5′-CGGCUGGAGCUGUUUGAGATT-3′ and 5′-GGAGUGAUUUUU CUAUCGGTT-3′
Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).
Incubate for 5 minutes at room temperature and add to cells.
Replace medium after 5h and re- incubate cells for 2 days at 37°C. |
Publication protocol
The HT1080 cells were transiently transfected with the siRNA oligonucleotides using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's instructions. Starved HT1080 cells were transfected with 20 nM siRNAs for 5 h and stabilized for 48 hr in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in 5% CO2 for further assays. The siRNA oligonucleotide sequences for ICAM-1 are as follows: 5′-CGGCUGGAGCUGUUUGAGATT-3′ and 5′-GGAGUGAUUUUU CUAUCGGTT-3′ (Invitrogen).
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