FIGURE 3.

RLIP76 levels in normal breast cells versus breast cancer cells and effect of 2HF and RLIP76 antisense on protein expression levels of RLIP76. A, 100 μg of protein from normal (MCF10a) and malignant (MCF7, SKBR3, MDAMB231, T47D, and TMD231) cells were loaded on the gel and blots were probed with anti-RLIP76 IgG. β-actin was used as a loading control. The experiment was performed three times and the image show one representative experiment. B, Breast cancer cells were treated with vehicle control (C) and 25 μM 2HF (T) for 24 h for Western blot analyses. Western blots was also done 24 h after treatment with scrambled (C) or RLIP76 antisense (T; 20 μg/mL final concentration), using Maxfect transfection reagent. Fifty microgram of protein were loaded on the gel, and blots were probed with anti-RLIP76 IgG. β-actin was used as a loading control. Numbers below the blots represent the fold change in the levels of proteins as compared to Control as determined by densitometry. The experiment was performed three times and the image shown is representative of one experiment. Bar represent densitometry analysis