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. 2020 May 1;11:2141. doi: 10.1038/s41467-020-16030-0

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© The Author(s) 2020

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 — a Schematic representation of AAV-RAM-Tet-off-PA-Cre 3.0 application for targeting active neurons in mouse dorsal raphe nucleus using RAM element and optic cannula. b Experimental time course of AAV-RAM-Tet-off-PA-Cre 3.0 application in mouse dorsal raphe nucleus with restraint, which can activate c-fos, and BL illumination. The mouse groups were kept in doxycycline-free condition. c Representative immunofluorescent images of red fluorescent protein, mCherry (red) and neuronal activity marker, c-fos (green), in mouse dorsal raphe nucleus infected with AAV-RAM-Tet-off-PA-Cre 3.0 viruses in dark (n = 2), and BL illumination (n = 3). Optic cannulas are shown in yellow dashed lines. White arrowheads show c-fos- and mCherry-positive neurons in the area. Scale bar, 100 μm.