Fig.1

Validation of RAGE-overexpressing BC cells. A Kaplan–Meier plot showing the association of RAGE mRNA expression with the progression free interval (PFI) of the TCGA ER-positive BC patients. Samples were divided into RAGE high and low groups using the optimum cut-off. B-C Morphological appearance of wild type (MCF7/wt and T47D/wt) and RAGE-overexpressing (MCF7/RAGE and T47D/RAGE) cells in phase-contrast microscopy; scale bar: 250 μm. Enlarged details are shown in the separate boxes. Flow cytometric histograms in RAGE-overexpressing compared with wild type MCF7 (D) and T47D (E) cells. FITC, fluorescein isothiocyanate. Side panels show the percentage of RAGE-positive cells. F-G Immunoblots of RAGE in wild type (MCF7/wt and T47D/wt) and RAGE-overexpressing (MCF7/RAGE and T47D/RAGE) cells. Side panels show densitometric analysis of the blots normalized to β-actin, which was used as a loading control. Values represent the mean ± SD of three independent experiments performed in triplicate. H-I Evaluation of RAGE protein expression (green signal) by immunofluorescence experiment in wild type (MCF7/wt and T47D/wt) and RAGE-overexpressing (MCF7/RAGE and T47D/RAGE) cells; nuclei were stained by DAPI (blue signal). The images shown represent 10 random fields from three independent experiments. Scale bar: 25 μm. Side panels represent corrected total cell fluorescence (CTCF), which was calculated on at least 10 pictures from each sample. (*) indicates p < 0.05; (**) indicates p < 0.01; (**) indicates p < 0.001