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. 2023 Feb 22;6:208. doi: 10.1038/s42003-023-04563-1

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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. 4 — a Left panel: Urinary albumin/creatinine ratio (U-ACR) in WT, Alb-hKL, and Pod-hKL, 0, 1, 3, and 7 days after injection of NTS. ^$$$$p < 0.0001 vs. Pod-hKL and ^@@@@p < 0.0001 vs. WT. Right panel: Blood urea nitrogen in the different groups without injury (control mice) or 7 days after injection of NTS (NTS mice). ^####p < 0.0001 vs. WT-control; ^@p < 0.05 vs. WT-NTS. b Representative images of PAS (left panels) and TEM (right panels) of kidney cortex from WT-control and WT, Alb-hKL, Pod-hKL 7 days after NTS injection. c Percentage of affected and crescentic glomeruli were quantified from 30 randomly selected glomeruli in the corresponding PAS staining. Tubular casts were assessed over the whole cross-section giving a score between 0, no cast to 5, >80% of the tubules are affected (n = 11 WT-control; n = 10 WT-NTS; n = 5 Alb-hKL-NTS; n = 11 Pod-hKL-NTS). Quantification of mean glomerular basement membrane (GBM) thickness and the number of slits per μm GBM (n = 5 WT-control; n = 10 WT-NTS; n = 6 Alb-hKL-NTS; n = 8 Pod-hKL-NTS). ^###p < 0.001, ^####p < 0.0001 vs. WT-control; ^@p < 0.05, ^@@p < 0.01 vs. WT-NTS and *p < 0.05, ***p < 0.001 vs. Alb-hKL-NTS. d Representative images of immunofluorescent stainings of Nephrin, WT-1, and α-SMA in the different groups. e Quantification of the number of WT-1 positive podocytes in glomeruli (left panel), glomerular area (middle panel), and WT-1 positive podocytes/glomerular area (right panel) (n = 7 WT-control; n = 9 WT-NTS; n = 5 Alb-hKL-NTS; n = 9 Pod-hKL-NTS). α-SMA positive area was counted per cortical cross section (10 sections per sample; n = 6 WT-control; n = 10 WT-NTS; n = 5 Alb-hKL-NTS; n = 9 Pod-hKL-NTS). ^##p < 0.01, ^###p < 0.001, ^####p < 0.0001 vs. WT-control; ^@p < 0.05 vs. WT vs. WT-NTS and *p < 0.05, ***p < 0.001, ****p < 0.0001 vs. Alb-hKL-NTS. Scale bars: 50 μm, 30 μm, 5 μm (b), 10 μm, 50 μm (d). Statistical significance between groups was determined using one-way or two-way repeated measures ANOVA followed by multiple comparisons.

Fig. 4 — a Left panel: Urinary albumin/creatinine ratio (U-ACR) in WT, Alb-hKL, and Pod-hKL, 0, 1, 3, and 7 days after injection of NTS. ^$$$$p < 0.0001 vs. Pod-hKL and ^@@@@p < 0.0001 vs. WT. Right panel: Blood urea nitrogen in the different groups without injury (control mice) or 7 days after injection of NTS (NTS mice). ^####p < 0.0001 vs. WT-control; ^@p < 0.05 vs. WT-NTS. b Representative images of PAS (left panels) and TEM (right panels) of kidney cortex from WT-control and WT, Alb-hKL, Pod-hKL 7 days after NTS injection. c Percentage of affected and crescentic glomeruli were quantified from 30 randomly selected glomeruli in the corresponding PAS staining. Tubular casts were assessed over the whole cross-section giving a score between 0, no cast to 5, >80% of the tubules are affected (n = 11 WT-control; n = 10 WT-NTS; n = 5 Alb-hKL-NTS; n = 11 Pod-hKL-NTS). Quantification of mean glomerular basement membrane (GBM) thickness and the number of slits per μm GBM (n = 5 WT-control; n = 10 WT-NTS; n = 6 Alb-hKL-NTS; n = 8 Pod-hKL-NTS). ^###p < 0.001, ^####p < 0.0001 vs. WT-control; ^@p < 0.05, ^@@p < 0.01 vs. WT-NTS and *p < 0.05, ***p < 0.001 vs. Alb-hKL-NTS. d Representative images of immunofluorescent stainings of Nephrin, WT-1, and α-SMA in the different groups. e Quantification of the number of WT-1 positive podocytes in glomeruli (left panel), glomerular area (middle panel), and WT-1 positive podocytes/glomerular area (right panel) (n = 7 WT-control; n = 9 WT-NTS; n = 5 Alb-hKL-NTS; n = 9 Pod-hKL-NTS). α-SMA positive area was counted per cortical cross section (10 sections per sample; n = 6 WT-control; n = 10 WT-NTS; n = 5 Alb-hKL-NTS; n = 9 Pod-hKL-NTS). ^##p < 0.01, ^###p < 0.001, ^####p < 0.0001 vs. WT-control; ^@p < 0.05 vs. WT vs. WT-NTS and *p < 0.05, ***p < 0.001, ****p < 0.0001 vs. Alb-hKL-NTS. Scale bars: 50 μm, 30 μm, 5 μm (b), 10 μm, 50 μm (d). Statistical significance between groups was determined using one-way or two-way repeated measures ANOVA followed by multiple comparisons.