siRNA SKIL

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 30000 cells/well	The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) SKIL siRNA concentration- 50 nM Add INTERFERin to siRNA duplexes and incubate for 10min at RT after homogenization. Remove culture medium from cells and add incubate mixture. Incubate the plate at 37°C for 24 to 72 h.

Upstream tips
Seed 30000 cells/well
Protocol tips
The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) SKIL siRNA concentration- 50 nM Add INTERFERin to siRNA duplexes and incubate for 10min at RT after homogenization. Remove culture medium from cells and add incubate mixture. Incubate the plate at 37°C for 24 to 72 h.

Publication protocol

The knockdown of SKIL expression was done using small interfering RNA (AM16708, ID 107695) from Ambion (Ambion, Austin, TX, USA). The siRNA sequences (5′-to-3′) were:

1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense)

2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense)

Silencer® Negative Control #1 siRNA (AM4611, Ambion, Austin, TX, USA) was used as a non-targeting control. Cells were seeded into 24-well plates (30000 cells/well) in four replicates and were transfected the following day with 50 nM SKIL siRNA and scrambled siRNA. INTERFERin TM (PolyPlus Transfection, Strasbourg, France) and Opti-MEM were used for cell transfection.

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Manufacturer protocol

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