Fig 1: Urinary uromodulin levels in Bartter type 2 patients. (a) Amino acid composition of ROMK, with mutations found in Bartter type 2 patients (yellow, missense; red, nonsense). TM: Transmembrane domain. (b) Box and whiskers plot of urinary uromodulin (UMOD) levels of Bartter type 2 patients, control relatives and a healthy reference population (age = 20, eGFR = 80). Median, 25th and 75th percentiles are indicated on the respective plot. (c) Representative Western blot analysis of UMOD in urine samples from Bartter type 2 patients and control individuals. Samples were loaded according to urinary creatinine. (d) Western blot analysis of UMOD in control and deglycosylated (PNGase F treatment) urine samples from Bartter type 2 patients and control relatives. Samples were denatured by heating and treated according to the manufacturer’s instructions. Full length blot images can be found in Supplementary Figure S2.
Fig 2: Histological characterization, transcript analysis and uromodulin processing in Kcnj1-/- mice. (a) Haematoxylin and eosin (H&E) staining of 12 weeks Kcnj1+/+ and Kcnj1-/- kidneys. Scale bars: low magnification - 2 mm, high magnification - 100 µm (b) Quantitative PCR analysis of tubular marker genes in Kcnj1-/- kidneys. Glom (Glomerulus): Nphs2 (Podocin); PT (Proximal Tubule): Slc5a2 - Sodium/glucose cotransporter 2 (SGLT2), Aqp1 - Aquaporin 1; TAL (Thick Ascending Limb): Slc12a1 - Na/K/Cl cotransporter 2 (NKCC2), Umod - Uromodulin; DCT (Distal Convoluted Tubule): Slc12a3 - Na/Cl cotransporter (NCC), Pvalb - Parvalbumin; CNT/CD (Connecting Tubule/Collecting Duct): Aqp2 - Aquaporin 2, Scnn1b - Sodium channel epithelial 1 beta subunit (ß-ENaC), Scnn1g - Sodium channel epithelial 1 gamma subunit (?-ENaC); Cldn16 - Claudin 16 and Clcnkb (Cl channel Kb) are localized in both TAL and DCT. (c) Representative Western blot analysis on whole kidney lysates of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. ß-actin was used as a loading control. (d) Representative Western blot analysis on kidney membrane fractions of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. Flotillin-1 was used as a loading control, whereas GAPDH was used to test cytosolic contamination. (e) Representative Western blot analysis on kidney cytosolic fractions of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. GAPDH was used as a loading control, whereas Flotillin-1 was used to test membrane contamination. (f) Immunofluorescence analysis for UMOD (green) and NKCC2 (red) in Kcnj1+/+ and Kcnj1-/- serial kidney sections. Scale bars: low magnification - 50 µm, high magnification - 15 µm. Full length blot images can be found in Supplementary Figure S2.
Supplier Page from Abcam for Mouse Uromodulin ELISA Kit