Protocol tips
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PPAR-delta siRNA concentration-40 pmol.
Incubate TransIT-TKO Reagent:siRNA complexe mixture at room temperature for 15–30 minutes to allow sufficient time for complexes to form and add to cells.
Incubate for 24–72 hours.
It is not necessary to replace the complete growth medium with fresh medium
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Protocol tips |
PPAR-delta siRNA concentration-40 pmol.
Incubate TransIT-TKO Reagent:siRNA complexe mixture at room temperature for 15–30 minutes to allow sufficient time for complexes to form and add to cells.
Incubate for 24–72 hours.
It is not necessary to replace the complete growth medium with fresh medium
|
Publication protocol
This method was previously reported [13]. In brief, duplexed RNA oligonucleotides for human PPAR-delta (siGENOME SMARTpool TM) were obtained from Thermo Fisher Scientific Inc., Taipei, Taiwan. DU145 cells were transfected with 40 pmol of PPAR-delta siRNA or scramble siRNA (scramble) using transfection reagents (TransIT-TKO, Mirus) as directed in the manufacturer’s protocols. The adequacy of PPAR-delta silencing was tested by immunoblot to optimize the experimental conditions (siRNA dose and time after transfection) according to previous reports [38–40].
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Manufacturer protocol
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