Figure 4.

Withaferin A (WA) administration inhibits activity of complex III in the tumor of mouse mammary tumor virus–neu mice. A) Activity of complex III in tumor lysates from mice of the WA treatment group and the control group. Complex III activity was measured using oxidized cytochrome c and ubiquinol as substrates. Results represent mean complex III activity (n = 10) with their 95% confidence intervals (error bars). B) Activity of complex IV in tumor lysates from mice of the WA treatment group and the control group. Complex IV activity was measured using reduced cytochrome c as the substrate. Results represent mean complex IV activity (n = 10) with their 95% confidence intervals (error bars). C) 8-Hydroxy-2′-deoxyguanosine (8-OHdG) level by immunohistochemistry in representative tumors of a mouse from the WA treatment group and the control group. Immunohistochemical staining was performed with anti-8-OHdG antibody and analyzed by nuclear algorithm using Aperio ImageScope software (×200 magnification, scale bars = 100 μm). Bar graph shows mean H-score for 8-OHdG (n = 7) with their 95% confidence intervals (error bars). D) 8-OHdG level by immunohistochemistry in representative normal mammary ducts of a mouse from the WA treatment group and the control group. Immunohistochemical staining was performed with anti-8-OHdG antibody and analyzed by nuclear algorithm using Aperio ImageScope software (×200 magnification, scale bars = 100 μm). Bar graph shows mean H-score for 8-OHdG (n = 7) with their 95% confidence intervals (error bars). Statistical significance of differences was determined by two-sided Student t test (A–D).