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. 2019 Oct 11;10:4635. doi: 10.1038/s41467-019-12554-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Blood clearance and brain deposition level of intravenously injected NLCs in mice. a Blood clearance of intravenously injected Cy5.5-CGY and Cy5.5 scrambled peptide (SP). b Brain levels of Cy5.5-CGY and its scrambled formulation after correction for the blood content of the brain. c Blood clearance of intravenously injected Cy5-siRNA/FAM-CGY complexes compared with naked Cy5-siRNA and CY5-siRNA complexion with FAM-scrambled peptide (SP). d Brain levels of Cy5-siRNA delivered through the aforementioned formulations in (c). The brain level of siRNA is subtracted from the organ blood level. The right section shows animal imaging of siRNA accumulation in the head region. A typical animal image for each formulation is shown. In all experiments the final concentration of the peptide conjugate was 10 µM and that of siRNA was 48 nM, based on blood volume calculation equivalent to 6% of the body weight. All injections were done in groups of three animals and each experiment was repeated three times. The results are mean values ± s.d. e Fluorescent microscopy images of brain sections (following transcardial perfusion and fixation) showing the presence of intact Cy5-siRNA/FAM-CGY in the brain (yellow colour in overlay). f Fluorescent microscopy images of the brain sections showing the presence of FAM-CGY in the brain and co-localisation with CD31⁺ endothelial cells as well as TfR and RAGE (some examples are depicted with arrows in overlay images). CC = cerebral cortex, Hc = hippocampus and * denotes deep cerebral white matter. g Fluorescent microscopy images of the brain sections showing the presence of Cy5-siRNA (delivered through complexion with FAM-CGY) in the brain and co-localisation with CD31 positive endothelial cells and RAGE (some examples are depicted with arrows in overlay images), but not TfR. In both (f) and (g) NLCs appear in the brain parenchyma. Each experiment was repeated at least three times (in three different animals) with different sections (n = 9). Scale bar = 100 μm. Source data are available in the Source Data file