Phd2 sirna

Arg-G4-siRNA was prepared by the simple commixture. Arg-G4 nanoparticles were dissolved in 50 mM Tris-HCl buffer (pH 7.4). PHD2 siRNA (Santa Cruz Biotechnology, Santa Cruz, Dallas, TX, USA), which was dissolved in DEPC (diethylpyrocarbonate) water, was dropwise added into the nanoparticle solution at various nanoparticle to DNA nitrogen-phosphorus (N/P) ratios (1, 5, 10, 15, and 20). At room temperature, the mixture was immediately vortexed gently for 20 minutes and then allowed to sediment for 10 minutes. The resulting Arg-G4-siRNA complexes were used for the following experiments freshly.

For the RNA interference assay, IL-32- and VHL-specific siRNA oligonucleotides were purchased from Samchully Pharmaceuticals (Seoul, Korea). The following sequences were used for the construction of the siRNAs: IL-32 siRNA forward 5'-GCUCUCUGUCAGAGCUCUU-3' and reverse 5'-AAGAGCUCUGACAGAGAGC-3'; and VHL siRNA forward 5'-CCACAGCUACCGAGUGUAUTT-3' and reverse 5'-AUACACUCGGUAGCUGUGGTT-3'. CHIP siRNA, Keap1 siRNA and PHD2 siRNA were purchased from Santa Cruz Biotechnology. PKCδ siRNA was purchased from Bioneer (Seoul, Korea). The cells were transfected with 10 nM of siRNA using RNAiMAX (Invitrogen, Carlsbad, CA, USA).