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. 2023 Jun 3;14:3236. doi: 10.1038/s41467-023-39050-y

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© The Author(s) 2023

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. 1 — a Schemes illustrating AA injury design, integrated data annotation strategy and renal cell types after dataset integration (UMAP plot). S3 type 2 PT cells (S3T2); Loop of Henle and principal cells (LOH/CD-PC); macrophages and dendritic cells (Macro/Dend.); S1 PT cells (S1); distal convoluted tubular, connecting and intercalated cells (DCT.CNT.CD-IC); myofibroblasts and stromal cells (Myo/St. mixed); S3 PT cells (S3); endothelial and glomerular cells (Endo/Glom). b Representative spatial plots of integrated clusters resolved on uninjured and injured kidney slices showing impaired cortico-medullary organization following injury. c Cell type proportions per conditions in the integrated data. d Representative H&E staining and spatial transcriptomics Cloupe browser kidney images with resolution of proximal tubule injury and fibrotic markers at baseline and 3 weeks after AA injury (one uninjured Tgfbr2^fl/fl kidney, one uninjured γGT-Cre;Tgfbr2^fl/fl kidney, one injured Tgfbr2^fl/fl kidney and one injured γGT-Cre;Tgfbr2^fl/fl kidney). Scale bars in source data. e Representative H&E images of kidneys from uninjured and 6 weeks AA injured mice. Arrows indicate injured tubules and interstitial cell infiltrate. f Relative Kim-1 mRNA levels in renal cortices 6 weeks after AA injury measured by RT-qPCR using Actb mRNA levels for normalization; n = 9 (Tgfbr2^fl/fl) and 8 (γGT-Cre;Tgfbr2^fl/fl) mice, p = 0.014. g Quantification of CD45+ cells from uninjured and injured renal leukocytes analyzed by FACS; n = 6 per genotypes of uninjured mice; n = 5 (Tgfbr2^fl/fl) and 7 (γGT-Cre;Tgfbr2^fl/fl) for injured mice, p < 0.0001. h Sirius red staining of kidneys from uninjured and AA injured mice showing collagen accumulation in red. Arrows indicate fibrotic areas. i Quantification of Sirius red positive area; n = 13 (Tgfbr2^fl/fl) and 17 (γGT-Cre;Tgfbr2^fl/fl) mice, p = 0.0174 (two-tailed Mann–Whitney test). j Plasma BUN levels measured 6 weeks after AA injury; n = 18 (Tgfbr2^fl/fl) and 13 (γGT-Cre;Tgfbr2^fl/fl) mice, p = 0.0175 (two-tailed Mann–Whitney test). All scale bars (e and h) represent 100 μm; dots represent the number of animals per group (f, g, i and j). Data are presented as mean values ± SEM. Statistical significance was determined by unpaired Student’s t test (two groups) or two-way ANOVA followed by Sidak’s multiple comparisons test with p < 0.05 considered statistically significant unless otherwise stated. *p < 0.05; ****p < 0.0001. Source data are provided as a Source data file.