Volume 66, Issue 5, Pages (November 2004)

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Volume 66, Issue 5, Pages 1859-1865 (November 2004) Heterozygous mice for TGF-βIIR gene are resistant to the progression of streptozotocin- induced diabetic nephropathy Hwal Woong Kim, Bong C.H.O. Kim, Chi Young Song, J.I. Hoon Kim, Hye Kyoung Hong, Hyun Soon Lee Kidney International Volume 66, Issue 5, Pages 1859-1865 (November 2004) DOI: 10.1111/j.1523-1755.2004.00959.x Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 1 Light microscopic views. Shown are glomeruli from wild-type control mouse (A), transforming growth factor-β (TGF-β) type II receptor (TGF-βIIR) gene heterozygous (TGF-βIIR+/−) (HT) control mouse (B), wild-type diabetic mouse (C), and HT diabetic mouse (D) at 28 weeks of experiment. Periodic acid-Schiff (PAS) stain, ×400. Kidney International 2004 66, 1859-1865DOI: (10.1111/j.1523-1755.2004.00959.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 2 Immunoperoxidase staining. Shown are transforming growth factor-β (TGF-β) type II receptor (TGF-βIIR) (A to D) and phosphorylated Smad2/Smad3 (E to H) in the glomeruli of the wild-type controls (A and E), HT controls (B and F), wild-type diabetic mice (C and G), and HT diabetic mice (D and H). In wild-type (A and E) and HT (B and F) controls, there is no staining for TGF-βIIR (A and B) and phosphorylated Smad2/Smad3 (E and F). In the wild-type diabetic mice, glomerular cells exhibit positive staining for TGF-βIIR (arrows) (C) and phosphorylated Smad2/Smad3 (G). In the HT diabetic mice, there is no staining for TGF-βIIR (D), but a few glomerular cells exhibit positive staining for phosphorylated Smad2/Smad3 (H). Kidney International 2004 66, 1859-1865DOI: (10.1111/j.1523-1755.2004.00959.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 3 Nuclear binding activity to CAGA sequence by Southwestern histochemistry. The glomeruli of the wild-type control (A), HT control (B), wild-type diabetic mouse (C), and HT diabetic mouse (D). An increased amount of nuclear proteins that bind to a labeled CAGA sequence is evident in the glomerular cells of wild-type diabetic mouse (C) compared with wild-type control (A), HT control (B), and HT diabetic mouse (D). For competition experiments, sections from wild-type diabetic mice were preincubated with a 100-fold excess of unlabeled oligonucleotide probe before addition of labeled probe (E). Kidney International 2004 66, 1859-1865DOI: (10.1111/j.1523-1755.2004.00959.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 4 Northern blot analysis of α1(IV) collagen mRNA in the kidney tissue. Shown are wild-type control (WT-CON) (lane 1), wild-type diabetic mice (WT-DM) (lane 2), HT control (HT-CON) (lane 3), and HT diabetic mice (HT-DM) (lane 4). The blots were hybridized with [32P]-labeled cDNA for α1(IV) collagen (A) and β-actin (B). (C) Quantitative expression of α1(IV) collagen mRNA abundance after correcting for the β-actin signal. The α1(IV) collagen mRNA levels of experimental mice are expressed as percent increases compared with the mRNA levels of wild-type controls. Values are means ± SD. *P < 0.05 vs. HT diabetic mice, wild-type control, or HT control. Kidney International 2004 66, 1859-1865DOI: (10.1111/j.1523-1755.2004.00959.x) Copyright © 2004 International Society of Nephrology Terms and Conditions