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. 2017 Oct 25;7:14058. doi: 10.1038/s41598-017-14565-9

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© The Author(s) 2017

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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Decreased association between BK_Ca α1 and BK_Ca β1 subunits in vascular smooth muscle cells from diabetic patients. (A–C) Differential interference contrast (DIC) and confocal fluorescent PLA puncta (red) and DAPI (blue) images of freshly dissociated human vascular smooth muscle labeled with mouse-anti BK_Ca α1 (A), rabbit-anti BK_Ca α1 (B), and rabbit-anti BK_Ca β1 (C) antibodies. (D,E) DIC (left) and fluorescence PLA (red)/DAPI (blue) (right) images of dissociated vascular smooth muscle from non-diabetic (D) and diabetic (E) patients co-labeled with two distinct antibodies for the BK_Ca α1 subunit. (F,G) DIC (left) and fluorescence PLA (red)/DAPI (blue) (right) images of dissociated human vascular smooth muscle from non-diabetic (F) and diabetic (G) patients co-labeled for BK_Ca α1 and BK_Ca β1 subunits. Scale bar = 10 μm. (H) Quantification of PLA fluorescent puncta per μm² cell area for non-diabetic and diabetic vascular smooth muscle cells labeled for mouse-anti BK_Ca α1 (n = 19 cells), rabbit-anti BK_Ca α1 (n = 27 cells), rabbit-anti BK_Ca β1 (n = 12 cells), mouse-anti BK_Ca α1 + rabbit-anti BK_Ca α1 (n = 24 non-diabetic, 14 diabetic cells); mouse-anti BK_Ca α1 + rabbit-anti BK_Ca β1 (n = 27 non-diabetic, 24 diabetic cells). *P < 0.05, Mann-Whitney test. Significance for columns 1, 2, 3 and 5 was compared to column 4, and column 7 was compared to column 6.