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. 2024 Sep 9;25(10):4387–4409. doi: 10.1038/s44319-024-00244-0

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A–E) Representative confocal images of immunofluorescence performed on the WT mouse brain injected with either LV_EGFP (A-A”’ and D-D”’), LV_NPY (B-B”’), or LV_Y2-NPY (C-C”’ and E-E”’). The green signal highlights the mossy fiber pathway of transduced dentate granule cells projecting their axons onto CA3 pyramidal neurons. (A–C) Staining for DAPI (blue), EGFP (green), and NPY (red) are shown. (D, E) Staining for DAPI (blue), EGFP (green), Y2R (red), are shown. Scale bar = 20 µm. (F, G) Representative western blot (F) and quantification (G) of the indicated proteins in extracts from hippocampi of WT mice injected with the indicated vectors. Protein levels are normalized for loading (Tubulin) and are shown as fold change over control (LV_ EGFP = 1 ± 0.05, n = 8 mice; LV_NPY = 1.66 ± 0.16, n = 7 mice; LV_Y2-NPY = 1.57 ± 0.22, n = 7 mice p = 0.0007). Values are presented as means ± SEM. Statistical significance was calculated using the Kruskal–Wallis one-way analysis of variance followed by Dunn’s post hoc test. *p < 0.05; **p < 0.01. (H) Representative western blot of the indicated proteins in the membrane fraction (Mem) or total extract (Lys) of hippocampi from WT mice injected with the indicated vectors. The arrow indicates the Y2-flag band in the western blot image. pcl pyramidal cell layer, mf mossy fiber. Source data are available online for this figure.