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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Neurobiol Dis. 2019 Mar 5;127:163–177. doi: 10.1016/j.nbd.2019.03.001

Figure 1. Characteristics of a non-viral peptide derived from ApoB that delivers a short RNA targeting α-syn to neuronal cells.

Figure 1.

(A) Diagrammatic illustration of the original 38-amino acid peptide derived from the Apolipoprotein B protein for delivery of proteins and peptides for receptor mediated BBB transcytosis to the CNS and the novel shorter, 11 amino acid (underlined) fragment coupled to a 5 Glycine linker and a 9 Arginine positively charged tail utilized for delivering siRNA into neuronal cells. (B) The siRNA targeting α-syn was incubated with different molar ratios of the ApoB11 peptide for 30 minutes. Complexed RNA and protein were run through an agarose gel and unconjugated RNA was observed by ethidium bromide staining. (C, E) Immunocytochemical and laser scanning microscopy analysis of N2A mouse neuronal cells incubated with FITC labeled siRNA targeting α-syn added at a ratio of 1:40 siRNA:C2-9r (derived from rhabdovirus positive control) or 1:10 siRNA:ApoB11 (respectively) and analyzed at different time points for uptake of the FITC labeled siRNA. Neuronal cells were labeled with an antibody against MAP2 and visualized with Tyramide Red. (D, F) Fluorescence levels representative of FITC tagged siRNA:C2-9r or siRNA:ApoB11 (respectively) trafficking into neuronal cells were estimated after 3, 24 and 48 hrs post incubation and expressed as pixel intensity. Levels rapidly increased and plateau after 3 hrs. Scale bar represents 15μm.