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. 2018 Jan 11;8:545. doi: 10.1038/s41598-017-19071-6

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© The Author(s) 2018

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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Verification of caveolin-1 deficiency. (a,b) Representative confocal images of control WT (a; n = 4) and Cav1-deficient kidneys (b; n = 4) after double immunofluorescence staining for Cav1 (green signal) and AQP2 (red signal) showing strong basolateral Cav1 labeling in WT CD principal cells identified by positive luminal AQP2 signals (exemplified by arrows); no corresponding Cav1 signal was detected in Cav1−/− kidneys (arrows). (c,d) Double staining for vascular Cav1 signal (green; arterioles marked with asterisks) and endothelial marker CD31 (red) in WT (c) and Cav1−/− kidneys (d); note strong Cav1 signal in WT but not in Cav1−/− vessels. (e–h) Representative transmission electron microscopy images of WT (e,g) and Cav1-deficient kidneys (f,h) documenting caveolae (arrows) in the plasma membrane of WT vascular smooth muscle (e) and CD principal cells (g), and absence of caveolae in Cav1-deficient vascular smooth muscle (f) and CD principal cells (h). (i,j) Pre-embedding labeling of WT and Cav1−/− kidney sections for Cav1 detected by transmission electron microscopy shows basolateral Cav1 labeling of principal CD cells in WT (arrows) but not in Cav1 kidneys.