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. 2021 Jan 22;12:549. doi: 10.1038/s41467-020-20832-7

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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 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. 8 — a–i Fetuin-A deficiency promotes accumulation of calcium mineral particles in hypoxic fetal kidneys. Calcium biominerals were detected by ATTO 488 fluorescently labeled fetuin-A (488-FA). Compared to normoxic or hypoxic wt mice, hypoxic Ahsg KO mice exhibited the strongest 488-FA staining intensity in the proximal tubulus (PT), indicative of an increased mineralized matrix turnover (a–c). Arrowheads in f and i point towards granular staining pattern in the papilla and cortex, respectively, reflecting bulk accumulation of 488-FA in kidneys of hypoxic Ahsg KOs. Such granules were not detectable in wt samples (d, e, g, h). j, k Ischemia-reperfusion injury (IRI) induces calcium mineral particles in adult kidneys. A granular staining pattern indicative of bulk accumulation of 488-FA at sites of calcium deposits was only present in IRI kidneys (k), but not in controls (j). l, m Fetuin-A supplementation reduced the expression of the fibrotic markers Col1a1 (l) and Col3a1 (m) in IRI kidneys 5 days after injury. No effect is seen in mice treated with physiological saline solution (NaCl). Data were analyzed from N = kidneys and are presented as mean ± SEM. Ordinary one-way ANOVA with Dunnett’s multiple comparisons test. Individual P-values are denoted above the comparison lines. (****P < 0.0001; ***P < 0.001). Images are representative of at least three independent experiments (a–k). Scale bar = 100 µm (a–k). Source data are provided as a Source Data file.