Figure 7
Tumor volumes (panel A) and the number of metastases per lung after mastectomy (panel B) in immunodeficient NSG mice injected orthotopically with human MDA-MB-436 triple negative BC cells and treated with Met, Phe, Asp, At, Met and Asp, Met and At vs controls.
Representative vessels in tumors of immunocompetent FVB mice injected with murine MMTV-Erbb2 BC cells (panel C). Using immunofluorescence and confocal microscopy, z-stack images depicted CD31+ endothelial cells (green), alpha-SMA+ pericytes (red) and DNA (DAPI, blue). Squares are representative of an area of 775 × 775 μm3. Panel D shows the relative frequency of microvessel density (MVD), alphaSMA+ and CD31+ cells in the tumors of immunocompetent FVB mice injected with murine MMTV-Erbb2 BC cells (panel C). Results observed in immunodeficient NSG mice injected with murine MMTV-Erbb2 BC cells or human MDA-MB- 436 BC cells were not significantly different (data not shown). Panel E shows AMPK activation in MDA-MB-436-derived tumors generated in immunodeficient NSG mice injected with human MDA-MB-436 BC cells and treated with Met or Phen alone or in combination with Aspirin (Asp) and/or Atenolol (At). Panel F shows mTOR activation in the same tumors analyzed in A. Panel G shows Activation of 4E-BP1 in the same tumors analyzed in A. Panel H shows p70 activation in the same tumors analyzed in A. For each blot, the levels of phosphorylated protein over total protein were quantified by image-j, compared to those of control and represented by numbers below the bands. Actin was used as loading control. *P < 0.05, **P < 0.01, ***P > 0.005 vs controls.
