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. 2021 Oct 15;11:20556. doi: 10.1038/s41598-021-00159-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Hemosiderin accumulation and CD68-positive cells in the interstitium of podocyte-specific Crb2 knockout mice at 6 months of age and electron microscopic study. (a) The blue area was significantly larger in Crb2^fl/flpod-Cre^Tg/+ mice than in Crb2^+/+pod-Cre^Tg/+ or Crb2^fl/fl mice. Scale bars 100 μm. (b) The brown cells in the interstitium of Crb2^fl/flpod-Cre^Tg/+ mice were positive for CD68, suggesting that they were macrophages. CD68-positive cells were significantly more frequent in the interstitium of Crb2^fl/flpod-Cre^Tg/+ mice than in that of Crb2^+/+pod-Cre^Tg/+ or Crb2^fl/fl mice. HPF high-power field. Scale bars 50 μm. Data are presented as mean ± SD. Statistical analysis by one-way analysis of variance, followed by Scheffe's multiple comparison test; n = 5 in each group. ***P < 0.001, ****P < 0.0001. (c) Transmission electron microscopy showed foot process effacement in Crb2^fl/flpod-Cre^Tg/+ mice compared to Crb2^fl/fl mice. Scale bars 1 μm. The mean width of the foot processes in Crb2^fl/flpod-Cre^Tg/+ mice was significantly greater than that in Crb2^fl/fl mice. Data are presented as mean ± SD. Statistical analysis by Student's t-test; n = 3 in each group. **P < 0.01. (d) Scanning electron microscopy showed damage to the foot processes' urinary sides in Crb2^fl/flpod-Cre^Tg/+ mice compared to Crb2^fl/fl mice. Scale bars 1 μm. (e) Hemosiderin-laden macrophages were observed in the interstitial area of Crb2^fl/flpod-Cre^Tg/+ mice, but not in that of Crb2^fl/fl mice, at 6 months of age. Scale bars 10 μm.