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. 2017 Oct 19;2(10):6939–6957. doi: 10.1021/acsomega.7b00741

Figure 7.

Figure 7

Figure 7

Effect of incorporation of DOPE in the siRNA complexes on siRNA intracellular delivery. The cellular uptake studies were repeated with selected peptides to investigate the effect of addition of an additional lipophilic component, and the results are presented as the average of quantified fluorescence (A) and percentage of the cells positive for FAM-labeled siRNA uptake (B). Addition of DOPE to the siRNA complexes enhanced intracellular delivery of siRNA significantly for most of the selected peptides. Data are presented as mean, n = 3, and the error bars represent standard deviation. The asterisks indicate significant difference in cellular internalization as a result of DOPE incorporation into each complex. DOPE (low) and (high) indicate the siRNA uptake of the cells exposed to complexes of siRNA and DOPE (with no peptide) using the lowest (for complexes with peptide/siRNA w/w ratio of 20:1 and the highest (for complexes with peptide/siRNA w/w ratio of 120:1) DOPE final concentration used in the other study groups included in the experiment. (C) Samples of flow cytometry results for siRNA uptake: the uppermost row represents the fluorescence pattern of cells transfected with linear peptide/siRNA w/w ratio of 20:1 or 40:1, with or without DOPE incorporated into the carrier. The middle and bottom rows represent samples of fluorescence pattern of cells transfected with cyclic peptide/siRNA w/w ratio of 80:1 or 120:1, with or without DOPE. The gate P2 represents fluorescence of cells considered siRNA-free, whereas the cells in the P3 region are positive for siRNA uptake. (D) Fluorescent microscope images showing individual channels (green, blue, and red), and an overlay for each treatment group as well as “No treatment” cells as the negative control.