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. 2008 Jun 17;295(2):E456–E462. doi: 10.1152/ajpendo.00015.2008

Fig. 4.

Fig. 4.

Protein oxidation and immunofluorescence detection of protein radical formation after 5 days of acetone treatment. Mice were receiving 2% acetone in their drinking water for 5 days together with the ip administration of 20 μl 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) 3 times/day. A: incubation of liver slices with primary anti-DMPO and Alexafluor 488 secondary rabbit antibody shows enhanced protein oxidation in acetone-treated liver (intense green staining). Omitting the primary antibody or applying the anti-DMPO antibody only without DMPO treatment demonstrated only minor background fluorescence (data not shown). B: compared with wild-type mice treated with acetone, iNOS knockout mice show a significant decrease in protein radical formation, whereas no changes were detected in the liver of NADPH oxidase knockout mice. C: quantitative comparison of the levels of protein oxidation. Confocal images were compared using the Scion Image program, and the degree of protein oxidation was expressed as mean fluorescence intensity in each group. P < 0.05 vs. control group (*) and vs. acetone-treated group (#).