Figure 2
From: Higher Order Architecture of Designer Peptides Forms Bioinspired 10 nm siRNA Delivery System
Cellular uptake by 3T3 Swiss mice fibroblasts. (a) Confocal microscopy image (merged all channels (DIC, FITC, AF647)) of 3T3 cells incubated with V1/siRNA(AF647), V2/siRNA(AF647), and lipofectamine(LF)/siRNA. The dye associated with V1 (green), and the dye associated with siRNA (red), are clearly separated, Pearson’s coefficient = 0.243; V2 (green) is colocalized with the siRNA (red) indicated by the yellow color, Pearson’s coefficient = 0.713 The bar represents 50 µm (first panel) and 10 µm (second panel); All controls in supplementary Fig. 4. (b) Multi-color flow cytometry analysis of V1/siRNA complex. Each quadrant indicates populations of cells expressing fluorescence in the respective channel: Untreated (grey) cells in Q4, V1N/siRNA(Cy3) in Cy3 channel (Q3), V1/siRNA in FITC channel (Q1), V1/siRNA(Cy3) in both Cy3 and FITC channel (Q2). (c) Multi-color flow cytometry analysis of V2/siRNA complex. Each quadrant indicates populations of cells expressing fluorescence in the respective channel: Untreated (grey) cells in Q4, V2N/siRNA(Cy3) in Cy3 channel (Q3), V2/siRNA in FITC channel (Q1), V2/siRNA(Cy3) in both Cy3 and FITC channel (Q2). (d) Flow cytometry: comparison of fluorescence intensity in the Cy3 channel of cells incubated with V1N/siRNA(Cy3) (blue), V2N/siRNA(Cy3) (purple), lipofectamine/siRNA(Cy3) (green), siRNA(Cy3) (orange), and untreated (red); The concentration of siRNA was 1 µM in all cases, except lipofectamine/siRNA(Cy3), 0.1 µm. The table in Fig. 3d lists the % cells that are Cy3+, representing uptake of siRNA; the percent of cells that uptake was measured using gated population (Supplementary Figs 5, 6).
