Figure 7.

Combined treatment with Th1 cytokines and statin drugs potentiates K-Ras delocalization from membrane. MDA-MB-231 human breast cancer cells were cultured with no additives (No Tx), treated with recombinant Th1 cytokines (TNF-α and IFN-γ, 10 ng/mL each), a statin drug (Simvastatin or Fluvastatin, 1 µM), or the combination of Th1 cytokines and a statin drug (“Simva + Cyto” or “Fluva + Cyto”) for 24 h. Membrane proteins were separated from cytosolic proteins and the distribution of the K-Ras protein was analyzed by Western blot. (A) Representative Western blot images and (B) corresponding semi-quantitative densitometry analysis of K-Ras normalized to β-actin loading control. Representative Western blot images of whole-cell K-Ras, AKT, ERK ½, and phospho-ERK ½ after (C) 24 and (D) 72 h of treatment. Semi-quantitative densitometry analysis of the (E) GAPDH-normalized total AKT, (F) GAPDH-normalized total ERK-1/2, and (G) GAPDH-normalized phosphorylated ERK-1/2. All Western blot experiments were repeated at least three times. Displayed graphical results in panels (E), (F), and (G) are from at least three trials +/− SEM. Letter designations in panel (E) represent Tukey’s HSD comparisons: treatments with the same letter designation are not statistically different; when letter designations differ between treatments, the p-value is less than 0.05.