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. Author manuscript; available in PMC: 2014 Nov 11.
Published in final edited form as: Biomacromolecules. 2013 Oct 16;14(11):3903–3915. doi: 10.1021/bm401026n

Figure 6.

Figure 6

Enhancement of siRNA delivery by polymer vehicles Tr4(77) and CD4(143) upon the addition of free polymer to solution. To understand the role of free polymer in gene down regulation, polyplexes formed at N/P = 8 were added to cells, then at time points of 0.5 h or 2.5 h after initial transfection, a solution containing free polymer equivalent to N/P=42 was added to the wells (brining effective polymer-siRNA ratio to N/P=50 after a time delay). For luciferase gene down regulation (graphs A and C), the cells were transfected with 200 μL/well of siRNA or siCon (at a final RNA concentration of 100 nM/well). For Cy5-siRNA cellular uptake (graphs B and D), the cells were transfected with 500 μL/well of Cy5-siRNA (at a final concentration 40 nM/well). (A) Luciferase gene down-regulation 48 h after transfection with CD4(143)-siRNA polyplexes. (B) Cellular uptake of Cy5-siRNA in the different polyplex formulations with CD4(143) 3 h after transfection. Data is reported as the percentage of live cells positive for Cy5. (C) Luciferase gene down-regulation 48 h after transfection with Tr4(77)-siRNA polyplexes. (D) Cellular uptake of Cy5-siRNA in the different polyplex formulations with Tr4(77) 3 h after transfection. Data is reported as the percentage of live cells positive for Cy5. Notation is as follows: hashed bar (lighter color) indicates treatment with polyplexes containing siRNA and the solid bar (darker color) indicates treatment with polyplexes containing siCon negative control. The numbers above the bars indicate percentage of gene down-regulation. [Note the * symbol indicates significantly different values as compared to the cells only control (p<0.05)].