Fig. 1.

The receptor for advanced glycation end products (RAGE) is over expressed in endometrial cancer (EC) and associated with reduced survival. RAGE expression was determined by immunohistochemistry in biopsies (n = 67) from healthy patients (a; n = 25) and patients with type I (b; n = 24) or type II (c; n = 18) EC. Biopsies were formalin fixed and paraffin embedded before sectioning and staining with α-RAGE antibody. Representative images were acquired on a Zeiss Axio Imager 2 microscope and analyzed using the ZEN 2012 image analysis software. Scale bars are 50 μm. RAGE expression (H-score) was conducted blind by three of the authors (NT, LM & DG) independently and the mean score for each slide used (d). Kaplan-Meier survival curves were constructed using Graph Pad PRISM 6 based on survival (months) following surgery (e). Within type II EC patients, time to disease recurrence following surgery (months) was monitored (f) and correlated with RAGE expression (g). Biodistribution studies were perfomed in nude athymic mice, which were dosed intravenously with anti-RAGE antibody conjugated to the fluorophore Alexa-750 (3 mg/kg) and sacrificed after either 24 h or 3 wks. Organs were harvested and homogenized and the fluorescence from the tissue slurry measured using a fluorescence microplate reader (Varioskan LUX, ThermoFisher) at wavelength 750 nM. Fluorescence was normalised using the weight of the tissue and values expressed as Fluorescence Intensity per gram of tissue (h & i). Data points for RAGE expression (H-score) represent individual patients (d). Data were analyzed by ANOVA and Dunnett’s pairwise multiple comparison test; values differ from healthy, ***p < 0.001, *p < 0.05