Measurement of tissue ROS in PRCP^gt/gt mice. (A-B) Studies on the expression of fluorescence in aorta (A) and kidney (B). Frozen aortas and kidneys from WT and PRCP^gt/gt (gt/gt) were stained with DHE. (A-B) Photographed on a Zeiss LSM510 confocal microscope aperture 40×/1.3 oil immersion. The relative DHE fluorescence was measured by morphometric analysis on 40× microscopic images using MetaMorph (Molecular Devices). Data are expressed as the number of fluorescent pixels per nuclei. Data are mean ± SEM of ≥ 9 samples on aorta and ≥ 4 samples on kidney. (C) Superoxide levels (O₂^.−) were measured from freshly harvested mouse aortas using the chemiluminescent probe lucigenin. Data are mean ± SEM ratio of ALU per LDH units from WT or PRCP^gt/gt aortic pieces. (D) Superoxide levels (O₂^.−) were determined from N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid buffer solubilized mouse kidneys using the chemiluminescent probe lucigenin. Data are mean ± SEM ratio of arbitrary lucigenin fluorescent units (ALU) per milligram of renal tissue from WT or PRCP^gt/gt. (E) Hydrogen peroxide levels (H₂O₂) were determined from harvested mouse kidneys using Amplex Red. Data are mean ± SEM ratio of AFU per milligram of dried renal tissue from WT or PRCP^gt/gt. (F) An immunoblot for eNOS was performed on renal lysates from WT and PRCP^gt/gt. The normalized relative ratio of renal uncoupled/coupled eNOS in lysates from 10 mice from WT and PRCP^gt/gt was compared by analysis of band intensity using densitometer scanning. Immunoblot of high molecular weight kininogen was used as a loading control. *P < .05.