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Volume 54, Issue 2, Pages (August 1998)

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1 Volume 54, Issue 2, Pages 464-472 (August 1998)
Rabbit distal convoluted tubule coexpresses NaCl cotransporter and 11β-hydroxysteroid dehydrogenase II mRNA  Heino Velázquez, Aniko Náray-Fejes-Tóth, Teresa Silva, Eleanor Andújar, Robert F. Reilly, Gary V. Desir, David H. Ellison  Kidney International  Volume 54, Issue 2, Pages (August 1998) DOI: /j x Copyright © 1998 International Society of Nephrology Terms and Conditions

2 Figure 1 Open reading frame and predicted amino acid sequence of rabbit form of NaCl cotransporter. Numbers at left indicate amino acid position; numbers at right indicate base position. Nucleotide sequence is consensus of overlapping PCR fragments as described in text, and an additional full length clone obtained using a proofreading enzyme (XL Taq Polymerase, Perkin Elmer). Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions

3 Figure 2 Single nephron RT/PCR experiment using NaCl cotransporter primers. Ethidium bromide-stained band of expected size is visible only in DCT lane. Markers are pBR 322, MspI digest. Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions

4 Figure 3 Southern blot of multiple SN-RT/PCR experiments using proximal tubule (PT), distal convoluted tubule (DCT), and cortical collecting duct (CCD) samples (N = 6) and NaCl cotransporter primers. Reagent blank was negative and positive control was positive. No proximal tubules were positive, whereas all DCT samples and some CCD samples were positive. Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions

5 Figure 4 Single nephron RT/PCR experiment using 11-β-HSD primers. Ethidium bromide-stained band of expected size was visible in DCT and in CCD lanes. Markers are pBR 322, MspI digest. Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions

6 Figure 5 Southern blot of multiple SN-RT/PCR experiments using PT (N = 7), DCT (N = 7) and CCD (N = 9) samples and 11-β-HSD primers. Reagent blank was negative and positive control was positive. No proximal tubules were positive, whereas all DCT and CCD samples were positive. Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions

7 Figure 6 Rate of conversion of 1nm 3H-labeled corticosterone to 11-dehydrocorticosterone by 0.5mm of dissected rabbit nephron segments. Values are means ± se, N = 3. Rates of conversion with vehicle and PT were not different and numerically equal to zero. Significant rates of conversion were observed in both DCT and CCD. Kidney International  , DOI: ( /j x) Copyright © 1998 International Society of Nephrology Terms and Conditions


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