Assessing Proliferation and Apoptosis in Kidney Tissue

Cell proliferation was assayed in paraffin-embedded kidney tissue by incorporation of 5-bromo-2-deoxyuridine (Roche Molecular Biochemicals, Mannheim, Germany), as previously described [29 (link)]. Pregnant mice received an intraperitoneal injection of BrdU (100 mg/g of body weight) 2 h prior to sacrifice. BrdU-positive cells were identified using an anti-BrdU peroxidase-conjugated antibody (Roche Molecular Biochemicals, Mannheim). Immunoreactivity was visualized using Aminoethyl Carbazole horseradish peroxidase chromogen/substrate solution (Vector, USA). Apoptosis was assessed in paraffin-embedded kidney tissue using the cell death detection kit (Roche Molecular Biochemicals, Mannheim, Germany) and visualized using 3,3’-Diaminobenzidine (DAB) substrate solution (Vector, USA).