Stk11 siRNA

Protocol tips

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	Concentration of siRNA- 0.5 µM. Incubate transfection mixture for 10 min at room temperature. Replace medium after four hours and incubate for additional 24 h.

Protocol tips
Concentration of siRNA- 0.5 µM. Incubate transfection mixture for 10 min at room temperature. Replace medium after four hours and incubate for additional 24 h.

Publication protocol

LKB1 siRNA (Stealth RNAi, Life Technologies) and control siRNA (Life Technologies) were used. The 3T3-L1 adipocytes or C2C12 myotubes cultured in 60-mm dishes (Becton Dickinson) were transfected with a 0.5 µM concentration of either siRNA by using the transfection reagent Lipofectamine 2000 (Life Technologies). Prior to transfection, 12.5 µl of siRNA and 5 µl of transfection reagent were diluted with 500 µl of serum-free medium in separate tubes at first, and then the diluted siRNA and Lipofectamine were mixed together. The mixtures were incubated for 10 min at room temperature and then added dropwise to each culture well. Four hours after transfection, the medium was exchanged for fresh medium. The cells were cultured for 24 h prior to the experiment. The efficiency of LKB1 silencing was monitored by both quantitative reverse transcription-PCR and Western blotting.

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