Mechanisms Mediating the Effects of γ-Tocotrienol When Used in Combination with PPARγ Agonists or Antagonists on MCF-7 and MDA-MB-231 Breast Cancer Cells
Figure 9
Western blot analysis of γ-tocotrienol and PPARγ antagonists (GW9662 and T0070907) when used alone or in combination with each other to determine protein levels of CBP p/H300, CBP C-20, and SRC-1 in (a) MCF-7 and (b) MDA-MB-231 cells. MCF-7 cells were initially plated at cells/100 mm culture plate and treated with control or treatment media containing either 2 μM γ-tocotrienol, 3.2 μM GW9662, or 3.2 μM T0070907 alone and in combination. MDA-MB-231 cells were plated in a similar manner and cells were treated with control or treatment media containing 3 μM γ-tocotrienol, 6.4 μM GW9662, or 6.4 μM T0070907 alone or in combination. All cells were fed fresh treatment media every other day for a 4-day incubation period. Afterwards, whole cell lysates were prepared from cells in each treatment group for subsequent separation by polyacrylamide gel electrophoresis (50 μg/lane) followed by Western blot analysis. Scanning densitometric analysis was performed on all blots done in triplicate and the integrated optical density of each band was normalized with corresponding β-actin, as shown in bar graphs below their respective Western blot images. Vertical bars in the graph indicate the normalized integrated optical density of bands visualized in each lane ± SEM. as compared with vehicle-treated controls.
