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. 2017 Sep 15;7:11696. doi: 10.1038/s41598-017-12064-5

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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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Increased bridging processes in Col4a3−/− glomeruli. (a) Representative image of Col4a3−/− glomeruli shows primary processes giving rise to many “intermediate” processes with more branching formations. These intermediate processes intercross with each other and form “bridge”-like structures between primary and foot processes. Scale bar, 500 nm. (b) Representative image of foot processes of Col4a3−/− glomeruli in TEM. Arrow indicates a bridging foot process. (c) The foot process (FP) height is significantly increased in Col4a3−/− mice (n = 108) compared to wild-type (WT) mice (n = 114) (**p < 0.01 by Student’s t-test). (d) HIM images of podocyte processes in wild-type (WT) (left panel) and Col4a3−/− (right panel) mice. White arrows represent the width of main branch from major processes. Pseudo-colored picture was obtained by processing Fig. 4a in Adobe Photoshop (right panel). The pseudo green color represents podocyte major process and foot processes from a podocyte, and the pseudo red color represents podocyte major process and foot processes from another podocyte. Pseudo purple color represents microprojections originating from podocytes. MP, major processes; P1, main branch from MP; P2, second branch; P3, third branch. Scale bars, 200 nm. (e) The width of main branch from podocyte major processes is significantly increased in Col4a3−/− (n = 18) compared to WT (n = 23) glomeruli (***p < 0.001 by Student’s t-test). (f) Representative TEM images of processes in Col4a3−/− (left panel, copied from Fig. 4b, for comparison), Cd2ap-knockout (ko) and lipopolysaccharide (LPS) treated mice (right panel). While bridging processes are observed in Col4a3−/− glomeruli, diffuse foot process effacement and flattened foot processes are detected without bridging processes in Cd2ap-ko and LPS-treated glomeruli. Scale bars, 200 nm.