Characterization of Mas receptor in the Bdkrb2^−/− mice. (A) The influence of losartan on the thrombosis time in the carotid artery Rose Bengal model in Bdkrb2^−/− mice. Untreated (n = 10) or losartan-treated (10 mg/kg per day in drinking water) (n = 5) Bdkrb2^+/+ or untreated (n = 10) or losartan-treated (n = 5) Bdkrb2^−/− mice were compared on the Rose Bengal assay for carotid artery thrombosis. In all panels, the values shown are mean ± SEM. (B) AngII levels in untreated Bdkrb2^+/+ (n = 4) and Bdkrb2^−/− (n = 5) or losartan-treated Bdkrb2^+/+ (n = 5) and Bdkrb2^−/− (n = 4). (C) ACE mRNA levels in untreated or losartan-treated Bdkrb2^+/+ and Bdkrb2^−/− (n = 4 in each group). (D) Ang-(1-7) concentration in Bdkrb2^+/+, Bdkrb2^−/−, losartan-treated Bdkrb2^+/+, and losartan-treated Bdkrb2^−/− (n > 8 in each group). (E) Mas mRNA levels in untreated or losartan-treated Bdkrb2^+/+ and Bdkrb2^−/− (n = 4 in each group). (F) Immunoblots for renal Mas and AT2R. Kidney lysates from Bdkrb2^+/+ and Bdkrb2^−/− with equal total protein amounts were immunoblotted with antibody to Mas, kininogen, AT2R, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH), respectively. Kininogen serves as the loading control for Mas because Mas and GAPDH migrate to the same position on the SDS-PAGE. The figure is a representative gel of 4 individual studies of 4 pairs of different renal lysates. (G) Ratio of receptor Mas or AT2R to loading control, respectively, in renal lysate (n = 4 in each group). All data shown are the mean ± SEM. (A–E) Analyzed by 1-way analysis of variance with a Bonferroni correction and were found to be significantly different. P values shown represent an analysis of difference between 2 groups.