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Volume 54, Issue 3, Pages 877-888 (September 1998) Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions Rie Marumo, Sakiko Kaizuma, Shoji Nogae, Masayuki Kanazawa, Tokihisa Kimura, Takao Saito, Sadayoshi Ito, Mitsunobu Matsubara Kidney International Volume 54, Issue 3, Pages 877-888 (September 1998) DOI: 10.1046/j.1523-1755.1998.00051.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Histological sections (trichrome stain) 10 through the midwall of the left ventricle. (A) Sham-operated. (B) Infarct of 40% in congestive heart failure. Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 Northern blots of rBSC1 and GAPDH mRNA. mRNA expression on renal outer medulla. Five micrograms of total RNA was used in each lane. (A) Comparison between normal control and dehydrated (N = 10; *P < 0.05). (B) Comparison between sham-operated and CHF (N = 5; **P < 0.05). Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 (A) Competitive PCR using total RNAs from the renal cortex. (B) The intensity ratio of mimic cDNA to intrinsic rBSC1 is plotted against the amount of mimic cDNA. The amount of rBSC1 mRNA in the sample was calculated from the equivalence point (mimic cDNA/intrinsic rBSC1 = 1.0). (C) The average of five such determinations (N = 5). *P < 0.01 in comparison with sham-operated rats. Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 (A) Competitive PCR using total RNAs from the renal cortex. (B) The intensity ratio of mimic cDNA to intrinsic rBSC1 is plotted against the amount of mimic cDNA. The amount of rBSC1 mRNA in the sample was calculated from the equivalence point (mimic cDNA/intrinsic rBSC1 = 1.0). (C) The average of five such determinations (N = 5). *P < 0.01 in comparison with sham-operated rats. Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 Co-localization of Tamm-Horsfall glycoprotein and rBSC1 mRNA detected by the constructed cRNA probe. (A) Staining with anti-Tamm-Horsfall glycoprotein antibody. (B) Staining with DIG-labeled BSC1 antisense cRNA. Keys (a) and (b) represent the same tubule. Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 In situ hybridization of the renal outer medulla. Inner stripe (A, B, and C) and outer stripe (D, E, and F) of outer medulla and renal cortex (G, H, and I). Control sham-operated (A, D, and G), dehydrated rats (B, E, and H), and rats with myocardial infarction (C, F, and I). Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 Immunoblots of renal outer medulla using a specific antibody against rBSC1. (A) Fifty micrograms of protein from the renal outer medulla was used in each lane. (B) Densimetric analysis of each lane. Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 7 Labeling of TAL by a specific antibody against rBSC1. Inner stripe (A, B, and C) and outer stripe (D, E, and F) of the renal outer medulla and renal cortex (G, H, and I). Control sham-operated (A, D, and G), dehydrated rats (B, E, and H), and rats with myocardial infarction (C, F, and I). Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 877-888DOI: (10. 1046/j. 1523-1755. 1998 Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 877-888DOI: (10. 1046/j. 1523-1755. 1998 Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 877-888DOI: (10. 1046/j. 1523-1755. 1998 Kidney International 1998 54, 877-888DOI: (10.1046/j.1523-1755.1998.00051.x) Copyright © 1998 International Society of Nephrology Terms and Conditions