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. 2020 Jan 24;11:512. doi: 10.1038/s41467-019-14039-8

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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. 6 — a Left to right: immunofluorescence image showing double staining of cNav1.5 (red) and Cx43 (green) in human heart 294050 cryosection with sinoatrial node (SAN) and right atrial (RA) regions (n = 8) separated by white dotted line; magnification of image to show the distribution of cNav1.5 in the RA vs. the SAN; bar graph showing fluorescence signal intensity in the human SAN vs. RA. b Left to right: immunofluorescence image showing double staining of nNav1.6 (red) and Cx43 (green) in human heart cryosection with SAN and RA regions (n = 7) separated by white dotted line; magnification of image to show the distribution of Nav1.6 in the RA vs. the SAN; bar graph showing fluorescence signal intensity in the human SAN vs. RA. c Left to right: cNav1.5 (red) and α-actinin (green; staining cardiomyocytes) dual staining; nNav1.6 (red) and α-actinin (green) dual staining confirm the cardiomyocyte-specific localization of cNav1.5 and nNav1.6. d Serial sections staining cNav1.5, nNav1.6, Cx43, and a nerve bundle labeled by anti-tyrosine hydroxylase (TH: staining sympathetic nerves) show that nNav1.6 and cNav1.5 are predominantly found in the myocytes relative to nerve bundles. All presented images a–d were collected from human heart 294050. e Left: representative immunoblotting bands for cNav1.5, α-actinin (marker of cardiomyocytes), and Cx43. Right: summary data of immunoblotting results of cNav1.5 protein distribution in the human SAN (n = 6) and RA (n = 6), compared with GAPDH (middle) and α-actinin (right), respectively. Cx43 connexin-43, GAPDH glyceraldehyde 3-phosphate dehydrogenase. Data were represented in mean ± SD. For immunostaining, analysis was done using lme4 and emmeans packages in R 3.4.4. Predictors included Heart (treated as random effect) and Condition (fixed effect). Pairwise tests between Condition levels were adjusted using Tukey’s method. Western blotting data analysis was done using two-sided t-test. Source data and uncropped versions of the western blot are provided as a Source Data file.