Fluorescein lotus tetragonolobus agglutinin lta

Pkd2-KO left kidney was perfused with cold PBS and 4% PFA prior to collection. All other mouse kidneys were collected, fixed in 10% formalin, dehydrated, and embedded in paraffin using a standard protocol. Samples were sectioned at 5 μm and subjected to hematoxylin and eosin staining. Cyst index (cystic area/total kidney section surface area) was performed using ImageJ analysis software. For immunofluorescence staining, the following antibodies and dilutions were used: anti-PS (Regulus Therapeutics Inc, 1:1000), anti-phosphohistone H3 (1:400, Sigma-Aldrich H0412), Biotinylated Dolichos biflorus agglutinin (DBA; Vector Laboratories, 1:500), and Fluorescein Lotus tetragonolobus agglutinin (LTA; Vector Laboratories, 1:200). Secondary antibodies were conjugated to Alexa Fluor 488 or Alexa Fluor 594 (Molecular Probes, 1:400).

Anti-EGFR and anti-phospho-EGFR antibodies were purchased from Cell Signaling Technology (Beverly, MA). Anti-ERK and anti-phospho-ERK antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). An antibody specific to heparin-binding epidermal growth factor-like growth factor (HB-EGF) was from R&D Systems (Minneapolis, MN). Anti-GPR40 antibodies were obtained from Epitomics (Burlingame, CA) and Santa Cruz Biotechnology (Santa Cruz, CA). Fluorescein Lotus tetragonolobus agglutinin (LTA) was from Vector Laboratories (Burlingame, CA). Anti-aquaporin-2 (AQP2) antibody was from Abcam (Cambridge, MA). Anti-Tamm-Horsfall protein (THP) antibody was from AbD Serotec (Raleigh, NC). Anti-calbindin-D28K antibody was from Sigma-Aldrich (St. Louis, MO). EETs were a generous gift from Jorge Capdevila (Vanderbilt University). Heparin-Sepharose CL-6B column was from Amersham Pharmacia Biotech (Piscataway, NJ). AG1478 was purchased from Calbiochem (San Diego, CA). CRM197 and all other chemicals were from Sigma-Aldrich (St. Louis, MO).