Volume 54, Issue 4, Pages 1052-1062 (October 1998) Renal expression of transforming growth factor-β inducible gene-h3 (βig-h3) in normal and diabetic rats1  Richard E. Gilbert, Jennifer L. Wilkinson-Berka, David W. Johnson, Alison Cox, Tina Soulis, Leonard L. Wu, Darren J. Kelly, George Jerums, Carol A. Pollock, Mark E. Cooper  Kidney International  Volume 54, Issue 4, Pages 1052-1062 (October 1998) DOI: 10.1046/j.1523-1755.1998.00081.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Northern blot of βig-h3, TGF-β1, α1 (IV) collagen and 18S in control and diabetic rat kidneys. Increased gene expression of βig-h3 and TGF-β1 is seen in kidneys of diabetic rats. 18S rRNA is similar in control and diabetic groups. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 Quantitation of kidney βig-h3, TGF-β1 and α1 (IV) collagen mRNA in control (□) and diabetic (▪). Data are means ± sem of the ratio of optical density of βig-h3, TGF-β1 and α1 (IV) col mRNA to that of 18S rRNA relative to control values (designated an arbitrary value of 1).*P = 0.05,†P < 0.01, diabetic versus control Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 Correlation between βig-h3 and TGF-β mRNA in control (○) and diabetic (•) rat kidneys. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 Northern blot of βig-h3 gene expression in cultured proximal tubular cells exposed to TGF-β1 at concentrations of 0 to 10ng/ml. βig-h3 mRNA increased in response to exogenous TGF-β1 in a dose-dependent fashion. 18S rRNA expression was unchanged. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 βig-h3 gene expression in cultured proximal tubular cells exposed to TGF-β1 at concentrations of 0 to 10ng/ml. Data are means ± se of the ratio of optical density of βig-h3 to that of 18S rRNA relative to control values (designated an arbitrary value of 1). *P < 0.05 versus control,†P < 0.05 versus 0.1ng/ml. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 βig-h3 gene expression in cultured proximal tubular cells exposed to EGF and IGF-I 100ng/ml. Data are means ± se of the ratio of optical density of βig-h3 to that of 18S rRNA relative to control values (designated an arbitrary value of 1). Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 7 In situ hybridization for βig-h3 mRNA in longitudinal sections of control (left panel) and diabetic (right panel) rat kidney localizing gene expression to the inner cortex and outer stripe of outer medulla. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 8 In situ hybridization photomicrographs of βig-h3 mRNA. In the cortex hybridization is present in the vascular component of the juxtaglomerular apparatus in control (A) and in greater abundance in diabetic (B) rat kidneys. Glomeruli, tubules and macula densa showed no βig-h3 expression. In both control (C) and diabetic (D) kidneys proximal tubules in the inner cortex but not the outer cortex (A, B) intensely expressed βig-h3. No specific hybridization was detected in sections exposed to βig-h3 sense probe (E, F). Magnification ×400. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 9 Immunohistochemistry of type IV collagen in tubulointerstitium of control (A) and diabetic rats (B). Increased immunostaining is present in kidneys from diabetic rats compared with control animals. Magnification ×400. Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 1052-1062DOI: (10. 1046/j. 1523-1755 Kidney International 1998 54, 1052-1062DOI: (10.1046/j.1523-1755.1998.00081.x) Copyright © 1998 International Society of Nephrology Terms and Conditions