Fig. 8.
Curcumin-mediated increase in oxidative damage in lung tissue. CCSP-rtTA/Ki-rasG12C bitransgenic mice were treated with 500 μg/ml of DOX in the drinking water. Two days after the initiation of DOX treatment, mice were kept on the normal diet or were fed diet supplemented with 4000 p.p.m. of curcumin. Nine days after the initiation of DOX treatment, the mice were euthanized and the lungs removed and homogenized in digestion buffer. Five microliters (0.13–2.2 mg) of supernatant were denatured and derivatized with 2,4-dinitrophenylhydrazine (DNPH), the derivatized proteins separated on 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis gels, transferred onto a nitrocellulose membrane and incubated with a primary antibody to either the DNP moiety of the protein or β-actin followed by a secondary horseradish peroxidase-conjugated antibody. The upper bands in the blot are non-specific binding that occurs when 2,4-dinitrophenylhydrazine-treated lysates are probed with the antibody to β-actin.