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. 2020 May 12;13:75. doi: 10.3389/fnmol.2020.00075

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Copyright © 2020 Tsuneoka and Funato.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Modified in situ HCR without proteinase K treatment. Vglut2 mRNAs were detected in the mouse medial preoptic area (A,C,E) and PVT (B,D,F) using Alexa647-conjugated hairpin DNA (#S41). Amplification was performed for 2 h. (A,B) Sections were pretreated with proteinase K. (C,D) Sections were not treated with proteinase K. Maternal behavior-induced c-Fos immunoreactivity that was diminished by proteinase K treatment before hybridization. Panels (E,F) correspond to dashed areas in panels (C,D). Arrowheads indicate double-positive cells of c-Fos protein and Vglut2 mRNA. LPOA, lateral preoptic area; MPOA, medial preoptic area; PN, parataenial nucleus; PVT, paraventricular thalamus. Scale bars: 200 μm (A,C), 100 μm (B,D) and 40 μm (E,F).