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Volume 57, Issue 4, Pages 1643-1650 (April 2000) Diminished adenylate cyclase activity and aquaporin 2 expression in acute renal failure rats Soo W. Kim, Youn S. Jeon, Jong Un Lee, Dae G. Kang, Hyun Kook, Kyu Y. Ahn, Sung Z. Kim, Kyung W. Cho, Nam H. Kim, Jin S. Han, Ki C. Choi, M.D. Kidney International Volume 57, Issue 4, Pages 1643-1650 (April 2000) DOI: 10.1046/j.1523-1755.2000.00008.x Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 1 Western blot analysis of aquaporin 2 (AQP2) in the kidney of control and acute renal failure (ARF)-2 day rats. Immunoblots of AQP2 in the cortex (A), outer medulla (B), and inner medulla (C). Each column represents the densitometric analysis, showing mean ± SEM of six rats. Symbols are: (□) control; () ARF. *P < 0.05; **P < 0.01 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 2 Differential centrifugation of total kidney extracts in control and ARF-2 day rats. Abundance of AQP2 in the high-density membrane fraction (A) and low-density fraction (B). The immunoblot was reacted with affinity-purified anti-AQP2. Each column represents the ratio of the high-density membrane fraction over low-density fraction (HD/LD), showing mean ± SEM of eight rats. *P < 0.05 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 3 Western blot analysis of AQP2 in the kidney of control and ARF-7 day rats. Immunoblots of AQP2 in the cortex (A), outer medulla (B), and inner medulla (C). Each column represents the densitometric analysis, showing mean ± SEM of six rats. Symbols are: (□) control; () ARF. *P < 0.05; **P < 0.01 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 4 Differential centrifugation of total kidney extracts in control and ARF-7 day rats. Abundance of AQP2 in the high-density (HD) membrane fraction (A) and low-density (LD) fraction (B). Each column represents the ratio of HD/LD, showing mean ± SEM of eight rats. *P < 0.05 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 5 Immunohistochemical localization of AQP2 in the outer medulla in control (A), ARF-2 day (B), and ARF-7 day (C) rats. AQP2 labeling is found in both the apical plasma membrane and cytoplasmic domains. In ARF, there was a marked decrease in AQP2 labeling. Magnification ×360. Publication of this figure in color was made possible by a grant from MSD Korea Ltd, Kwangju, Korea. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 6 cAMP production in response to arginine vasopressin (AVP) in the cortex (A), outer medulla (B), and inner medulla (C) in control and ARF-2 day rats. Symbols are: (○) control rats; (•) ARF-2 day rats. Each point represents the mean ± SEM of six experiments. *P < 0.05; **P < 0.01 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 7 cAMP production in response to forskolin and sodium fluoride in the outer medulla in the control and ARF-2 day rats. Symbols are: (○) control rats; (•) ARF-2 day rats. Each point represents mean ± SEM of five experiments. *P < 0.05; **P < 0.01 vs. control. Kidney International 2000 57, 1643-1650DOI: (10.1046/j.1523-1755.2000.00008.x) Copyright © 2000 International Society of Nephrology Terms and Conditions