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. 2023 Apr 17;14:2182. doi: 10.1038/s41467-023-37968-x

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© The Author(s) 2023

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. 2 — a AAV-EF1α-DIO-GCaMP6s was injected into the VP of ChAT-Cre mice. b A representative image of a coronal section containing GCaMP6 (green)-labeled neurons and ChAT- and SP-antibody-stained neurons and tissue. c Efficiency (green) (74 ± 3.20%, n = 12 section from six mice) and specificity (red) (93 ± 2.4%, n = 12 sections from six mice) of GCaMP6-labeling of ChAT neurons in the VP (n = 5 mice). d Representative raw traces showing the responses of GCaMP6-labeled (green) and eYFP-labeled (black) ChAT neurons in the VP to pain-like stimulation (pressure) on the contralateral and ipsilateral hind paws. Arrow, pain-like stimulation. e Heat maps showing the relative fluorescent signals from five GCaMP6 mice (n = 22 trials) and five eYFP mice (n = 25 trials) before (3 s), during (2 s), and after (7 s) the application of stimulation to the contralateral and ipsilateral hind paws. Each panel includes data from five mice (4–5 trials from each mouse). f The fluorescence intensities of GCaMP6 (green) and eYFP (black) were transformed into their z-scores (mean ± SEM). Each trace summarizes data from the same trials as this in (e). g The peak amplitudes of GCaMP6 (n = 22 trials from 5 GCaMP6 mice) and eYFP (n = 25 trials from five eYFP mice) responses to pain-like stimulation on hind paws. Contralateral: t = 7.91, df = 45, P < 0.001; ipsilateral: t = 7.38, df = 44, P < 0.001, two-tailed t-test. **P < 0.01.