Figure 3
From: Higher Order Architecture of Designer Peptides Forms Bioinspired 10 nm siRNA Delivery System
Functional characterization of peptide/siRNA complex. (a) GFP silencing in 3T3-GFP cells: siRNA (1 µM, except lipofectamine (LF) 0.1 µM, Supplementary Fig. 10). V1N/siRNA and V2N/siRNA treated samples were carried out in the presence (solid) and absence (pattern) of 5% FBS. Statistical differences between 3T3-GFP cell: treated (+serum) vs. untreated (•); treated with lipofectamine vs. treated with complexes (*). Increase of the peptide:siRNA ratio did not improve silencing (b) 7-AAD cell viability assay (flow cytometry) was performed on 3T3 cells untreated (black) or treated with either varying concentrations of V1N/siRNA (dark grey) or lipofectamine (0.25 µg:5pmol ratio) (light grey) complexes at fixed ratios after 24-hour incubation. Increase in mean fluorescence intensity indicates non-viable cells (c) Cytokine assays: HMDM were treated with LPS (positive control) V1, siRNA or V1/siRNA complex (1 µM siRNA) and secreted levels of inflammatory factors, TNF and IL-6, were measured by Luminex multiplex analysis. Statistical differences between untreated and treated cells were calculated by one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
