Gene expression profiling in the remnant kidney model of wild type and kinin B1 and B2 receptor knockout mice J.P. Schanstra, M. Bachvarova, E. Neau,

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Gene expression profiling in the remnant kidney model of wild type and kinin B1 and B2 receptor knockout mice J.P. Schanstra, M. Bachvarova, E. Neau, J.L. Bascands, D. Bachvarov Kidney International Volume 72, Issue 4, Pages 442-454 (August 2007) DOI: 10.1038/sj.ki.5002172 Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 1 BP measurements and biochemical parameters of ESRD progression in SNX-WT, SNX-B1R-KO, and SNX-B2R-KO mice 20 weeks after SNX. (a) BP measurements; (b) biochemical parameters. Serum creatinine increases (*P<0.0001) and urinary creatinine decreases (*P<0.001) 20 weeks after SNX in the different animals groups, versus respective controls; n=4–6 mice/group. A tendency of increased proteinuria was observed in SNX animals but it did not reach statistical significance. Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 2 Network analysis of dynamic gene expression in the 20-week remnant kidneys based on the twofold gene expression list. The top four networks were merged and are displayed graphically as nodes (genes/gene products) and edges (the biological relationships between the nodes). Intensity of the node color indicates the degree of upregulated (red) or downregulation (green). Nodes are displayed using various shapes that represent the functional class of the gene product (square, cytokine; vertical oval, transmembrane receptor; rectangle, nuclear receptor; diamond, enzyme; rhomboid, transporter; hexagon, translation factor; horizontal oval, transcription factor; and circle, other). Edges are displayed with various labels that describe the nature of the relationship between the nodes: — binding only; → acts on. The length of an edge reflects the evidence supporting that node-to-node relationship, in that edges supported by more articles from the literature are shorter. Dotted edges represent indirect interaction. Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 3 The Hnf4a signaling pathway. Figure legends are as described in Figure 2. Edge labels: A, activation; B, binding; E, expression; PP, protein–protein binding; RB, regulation of binding; LO, localization. Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 4 Comparisons of the common and distinct gene expression across the different experimental conditions using the Venn diagram analysis. (a) Comparison for upregulated genes. (b) Comparison for downregulated genes. (c) The different experimental conditions subject to comparison are indicated (see details in text). Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 5 Hierarchical clustering of 20-week remnant kidneys that discriminates between SNX-B1R-KO and SNX-B2R-KO kidney samples. A subset of candidate genes were initially obtained by filtering on signal intensity (twofold), retaining 306 genes. One-way analysis of variance parametric test (Welch t-test; variances not assumed equal; P≤0.05) further selected 65 genes. Clustering analysis based on the 65 gene list was performed using the standard Condition Tree algorithm provided in GeneSpring. The mean appears grey, whereas red signifies upregulation, and green signifies downregulation (see legend bar). B1R SNX kidney samples are indicated in grey; B2R SNX kidney samples are indicated in blue. Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 6 Histological analysis of remnant kidney tissues. Histological appearance of glomeruli in (a) SNX-WT mice, (b) SNX-B1R-KO mice, and (c) SNX-B2R-KO mice. Representative photographs of Masson trichrome-stained renal sections of (d) 20-week remnant kidney tissues of SNX-WT mice, (e) SNX-B1R-KO mice, and (f) SNX-B2R-KO mice. Arrows indicated accumulation of interstitial collagen (blue). Bars represent the quantitative analysis where data are expressed as the mean±s.d. *P<0.05 versus SNX-WT. Original magnification × 200. Histological analyses were performed on five mice/group. Kidney International 2007 72, 442-454DOI: (10.1038/sj.ki.5002172) Copyright © 2007 International Society of Nephrology Terms and Conditions