Volume 54, Issue 2, Pages (August 1998)

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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 464-472 (August 1998) DOI: 10.1046/j.1523-1755.1998.00036.x Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

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 1998 54, 464-472DOI: (10.1046/j.1523-1755.1998.00036.x) Copyright © 1998 International Society of Nephrology Terms and Conditions