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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Free Radic Biol Med. 2008 Aug 16;45(9):1340–1351. doi: 10.1016/j.freeradbiomed.2008.08.013

Figure 2.

Figure 2

Analysis of NADPH oxidase activity by measurement of O2∸ and H2O2 production in the membrane fraction. (A) Accumulation of 3-carboxyproxyl (CP) in the membrane fraction of RASMCs (0.1mg/ml) followed by an increase in the low-field component of EPR spectrum (insert). The rates of O2∸ and H2O2 production were 184 pmol/mg/min and 658 pmol/mg/min, correspondingly. The O2∸ independent H2O2 production was 474 pmol/mg/min; (B) Detection of O2∸ in the membrane preparation of RASMCs (0.2 mg/ml) using CMH or CPH. Addition of cytosol inhibited O2∸ detection similar to SOD-supplementation; (C) Measurement of O2∸ production by NADPH oxidase in homogenate and membrane fraction of RASMCs (0.1mg/ml) at various concentrations of CPH; (D) Detection of O2∸ in the homogenate of RASMCs (0.2mg/ml) using CMH, CPH and CPH + SOD-inhibitor DETC.