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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. 5 — Brain localization of NLCs and NLC-siRNA complexes and demonstration of pharmacological activity. a, b Fluorescent microscopy images of brain cerebral sections at 4 h after the administration of FAM-CGY NLCs (final concentration of 10 μM) (a) and FAM-CGY/Cy5-siRNA complexes (FAM-CGY = 10 μM, Cy5-siRNA = 48 nM) (b), respectively. Astrocytes, neurons and microglia were labelled with anti-GFAP, anti-Tuj1, anti-Iba1 antibodies, respectively and visualised with a secondary fluorescent antibody (Cy-3 channel). Inserts represent enlarged sections. Arrows indicate FAM-CGY NLC and FAM-CGY/Cy5-siRNA complexes. Cy3-Channel: astrocytes or neurons or microglial cells; FAM-Channel: FAM-CGY NLCs, Cy5-Channel: Cy5-siRNA. Cell nuclei are stained with DAPI. Scale bar = 40 μm. The images show NLC localization to neurons and microglial cells, but not astrocytes. c Western blot demonstration of suppression of β-secretase 1 (BACE1) expression in hippocampus following a single intravenous injection of FAM-CGY/BACE1 siRNA complexes. Silencing was compared with FAM-CGY/siControl injection. BACE1-specific siRNA alone or delivered with the FAM-CGY scrambled peptide 1 (FAM-CGY SP) did not induce silencing. BACE1 expression was measured 48 h after administration. The results represent mean values of three separate experiments ± s.d. *p < 0.05, non-paired two-sided student t-test compared with respective siControl (control siRNA). Source data are available in the Source Data file