Journal:

Article Title: Protein tyrosine kinase is expressed and regulates ROMK1 location in the cortical collecting duct

doi: 10.1152/ajprenal.00301.2003

Figure Lengend Snippet: Confocal images showing the colocalization of aquaporin-2 (AQP2; green) and c-Src (red) in the renal cortex (A) and OM (B) from rats on a normal-K diet. Bar = 10 µM or as indicated.

Article Snippet: Immunocytochemical staining The slides were washed with 1× PBS for 15 min and permeablized with 0.4% Triton dissolved in 1× PBS buffer containing 1% BSA and 0.1% lysine (pH 7.4) for 15 min. Kidney slices were blocked with 2% goat serum for 1 h at room temperature and then incubated with ROMK antibody (Alomone, Jerusalem, Israel), c-Src monoclonal antibody (Transduction Lab), aquaporin-2 (AQP2) antibody (Alomone), or Tamm-Horsfall glycoprotein (THP) antibody (ICN, Pharmaceutical, Aurora, OH) for 12 h at 4°C.

Techniques:

Journal:

Article Title: Protein tyrosine kinase is expressed and regulates ROMK1 location in the cortical collecting duct

doi: 10.1152/ajprenal.00301.2003

Figure Lengend Snippet: Magnified confocal image (×100) showing the double staining of AQP2 and c-Src in the renal cortex (A) and OM (B). Bar = 10 µM.

Article Snippet: Immunocytochemical staining The slides were washed with 1× PBS for 15 min and permeablized with 0.4% Triton dissolved in 1× PBS buffer containing 1% BSA and 0.1% lysine (pH 7.4) for 15 min. Kidney slices were blocked with 2% goat serum for 1 h at room temperature and then incubated with ROMK antibody (Alomone, Jerusalem, Israel), c-Src monoclonal antibody (Transduction Lab), aquaporin-2 (AQP2) antibody (Alomone), or Tamm-Horsfall glycoprotein (THP) antibody (ICN, Pharmaceutical, Aurora, OH) for 12 h at 4°C.

Techniques: Double Staining

Journal: Kidney360

Article Title: A Novel Human Distal Tubuloid-on-a-Chip Model for Investigating Sodium and Water Transport Mechanisms

doi: 10.34067/KID.0000000992

Figure Lengend Snippet: Expression profile of distal nephron markers in differentiated tubuloids. (A) Left: Schematic illustration of DNS highlighting the TAL of the loop of Henle (TAL; SLC12A1/NKCC2) and CD (SCNN1a/ENaC, AQP2) markers. Right: qPCR analysis showing segment-specific gene expression across four donors after differentiation (fold change relative to basal medium, log scale, Mean±SD). Samples from five chips per donor were pooled to create each replicate, with three independent samples per condition for each of the four donors. (B) Immunofluorescence analysis of tubuloid cultures (20× magnification). Upper panels show full-length tubules; lower panels show magnified regions (dotted boxes). Staining shows expression of ZO-1 (tight junctions), ECAD (distal tubule), NKCC2 (TAL), and AQP2 (CD), all counterstained with DNA (representative images, n =2–3). (C) Comparative gene expression analysis between 3D (OrganoPlate) and 2D (tubuloids grown on static 24-well plates) culture conditions across four donors (fold change, n =3). Comparisons made within each donor due to variability in baseline expression. All qPCR data were normalized to the housekeeping gene GAPDH and expressed as fold change relative to the corresponding 2D static culture condition for each donor. 2D, two-dimensional; CD, collecting duct; DCT, Distal convoluted tubule; DNS, distal nephron segments; ECAD, E-cadherin; ENaC, epithelial sodium channel; PT, proximal tubule; TAL, thick ascending limb.

Article Snippet: , AQP2 , Santa Cruz Biotechnology , Rabbit , — , Sc-515770 , 1:300.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining