Fig. 8

RAGE inhibition prevents colony and mammosphere formation induced by Ins. Evaluation of colony formation in MCF-7 cells treated with Ins (20 nM, 3 weeks), alone and in combination with FPS-ZM1 (10 μM) (A, B). Representative images showing mammosphere formation in MCF-7 cells treated with Ins (20 nM), alone and in combination with FPS-ZM1 (10 μM) for 5 days. Only spheres with a diameter > 50 μm were counted. Scale bar: 50 μm (C, D). mRNA expression of ALDH1A3 in MCF-7 cells treated with vehicle (−) or Ins (20 nM, 4 h) alone and in combination with FPS-ZM1 (10 μM, 8 h), as evaluated by qRT-PCR. Values are normalized to the 36B4 gene expression and shown as fold changes of mRNA expression induced by Ins compared to cells treated with vehicle (E). Evaluation of mammosphere formation in cells derived from surgical specimens obtained from BC patients. Cells were isolated and grown in suspension culture in the presence of Ins (10 nM), alone and in combination with FPS-ZM1 (10 μM) for 10 days (F). Only spheres with a diameter > 50 μm were counted (G). Data shown are the mean ± SEM of at least two independent experiments. (**) p < 0.01; (***) p < 0.001