Figure 6

MCD potentiates the effect of tamoxifen by altering Cav-1 levels and Akt/ERK signaling. (A) Cav-1 mRNA levels were determined by semi-quantitative RT-PCR. A375, B16F10 cells and tumor from mice treated with indicated concentration of tamoxifen along with MCD, were processed for RT-PCR. β-actin was used as a loading control. (B and C) A375 and B16F10 cells, respectively, were treated with indicated concentration of tamoxifen in- combination with MCD and cholesterol, and whole cell lysate were subjected to Western blotting and expression level of Cav-1, pAkt, Akt, pERK and, ERK was assessed. (D and E) Tumor bearing mice were exposed to normal diet and cholesterol supplementation respectively, were treated with indicated dose of MCD and tamoxifen, and tumor lysates were subjected to Western blotting and expression level of Cav-1, pAkt, Akt, pERK and, ERK was assessed. GAPDH served as a loading control. (F) Schematic diagram of proposed mechanisms of MCD potentiating the effect of tamoxifen and its abrogation by cholesterol supplementation.