Author: Dai, Zhi; Arévalo, Maria T; Li, Junwei; Zeng, Mingtao
Title: Addition of poly (propylene glycol) to multiblock copolymer to optimize siRNA delivery Document date: 2014_1_1
ID: yawwtyxg_27
Snippet: Gene silencing and intracellular translocation Neuro 2a/GFP cells were seeded in 12-well plates at 8 × 10 3 cells/well and cultured for 24 h. Then, cells were transfected with copolymer complexes containing GFP-siRNA at various N/P ratios. The final concentration of siRNA was 80 nM in each well. GFP-siRNA complexed with-Lipofectamine™ 2000 was used as a control. After a 4 h incubation period, the transfection medium was discarded and supplemen.....
Document: Gene silencing and intracellular translocation Neuro 2a/GFP cells were seeded in 12-well plates at 8 × 10 3 cells/well and cultured for 24 h. Then, cells were transfected with copolymer complexes containing GFP-siRNA at various N/P ratios. The final concentration of siRNA was 80 nM in each well. GFP-siRNA complexed with-Lipofectamine™ 2000 was used as a control. After a 4 h incubation period, the transfection medium was discarded and supplemented with fresh medium containing 10% FBS. The cells were incubated for another 24 h. The medium was removed, and the cells were washed twice with cold PBS and detached using 0.25% trypsin-EDTA. GFP expression was quantified via flow cytometry using a Gallios flow cytometer (Beckmann Coulter) and analyzed using FlowJo software (TreeStar). The parent Neuro 2a cell line not expressing GFP was used to control for background fluorescence.
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