Transfection of Cells with siRNA

Transfection with different concentrations of SSOs was performed with PepFect14 (Psyclo Peptide, Inc., Shanghai, China) (Ezzat et al., 2011 (link)) or Lipofectamine® 2000 (LF2000) (Life Technologies, Invitrogen, CA, USA) or OptiMEM (gymnotic delivery), which was formulated in OptiMEM for 15 min at room temperature according to the protocols of the manufacturer, and was then added to the cells cultured in DMEM + 10% FBS. The complexes were left in the culture for 24 h, after which the cells were harvested for RNA isolation or luciferase measurement. Transfection with 5 nM of Silencer Select siRNA® (Life Technologies, Carlsbad, CA, USA), including NCL s70420 and TCP1 s224715, was performed with LF2000, formulated in OptiMEM 15 min at room temperature, according to the protocols of the manufacturer, and was then added to the cells cultured in DMEM + 10% FBS. The complexes were left in the culture for 48 h, after which the cells were harvested for RNA isolation.