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. 2019 Aug 23;26(4):1376–1398. doi: 10.1038/s41380-019-0491-4

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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. 1 — Expression patterns and regulation of Plxnb2 mRNA and Plexin-B2 protein in brain regions involved in fear and fear memory. a–c Analysis of β-galactosidase expression in mice expressing the LacZ gene in the Plxnb2 locus (Plexin-B2-LacZ mice) across the CA1, CA3, and DG areas of hippocampus (a), anterior cingulate cortex (rACC; b) and in the basolateral amygdala (BLA) and central amygdaloid nucleus (CeA; c). Typical examples, and quantitative summary of densitometric magnitude of LacZ staining in mock-treated mice and at day 3 following fear conditioning via foot shock stimulation. In panel b, bar graphs represent mean signal density over unit area over the whole ROI (demarcated by black lines in the examples shown in panel b) as well as a separate analysis of LacZ staining signal intensity in the high and low expression level areas (medial and lateral areas, respectively; indicated by arrow and arrowhead, respectively), normalized on the number of cells present in the ROI; n = 3 mice/group; at least 3 sections were analyzed per mouse. d Analysis of Plxnb2 mRNA expression across the CA1 area of hippocampus (upper panels), rostral anterior cingulate cortex (rACC; middle panels) and in the central amygdala (CeA; lower panels) using RNAScope in situ hybridization (ISH) technique. Typical examples (images) and quantitative summary (bar graphs) of signals achieved with the Plxnb2 probe in mock-treated mice and mice with fear conditioning at day 3 following foot-shock; bar graphs on the left represent the integral signal intensity normalized on the number of cell nuclei, whereas bar graphs on the right represent the mean number of signal dots per nucleus; n = 3 mice per group; at least 3 sections were analyzed per mouse. e Examples and densitometric quantification of signals in Western blot analysis of Plexin-B2 protein expression in lysates of CA1 area of hippocampus (upper panels), rACC (middle panels) and the amygdalae-enriched tissue (lower panels) in mock-treated mice and following fear conditioning at day 3 following foot-shock; data are represented as fold changes of the ratio of Plexin-B2 levels over loading control signal (β-Tubulin III); n = 4 mice per group. Student’s t-test. P < 0.05 indicated by * as compared to the corresponding control (mock) group. Error bars represent SEM. Scale bars represent 100 µm (a–c) and 5 µm (d)